text: University of British Columbia

characteristic: INSDC last update

text: live

text: Generic

text: A mountain pine beetle-attacked Lodgepole pine (Pinus contorta) tree was felled on Nov. 11, 2006 along the KayKay Forest Service Road northwest of Prince George, BC, Canada (approx. N 54 02.731' W 123 19.109') and the bolts stored at room temperature with waxed sawn ends to reduce dehydration until use. On Dec. 8, 2006, 13 live larvae of mixed instar and of undetermined sex were manually removed from the bolt and immediately placed whole in liquid nitrogen. An un-normalized, full-length enriched, directionally cloned library was prepared with the Creator SMART cDNA Library Construction Kit (Clontech, Mountain View, CA, USA) and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, Moscow, Russia). The Evrogen kit was used for some of the protocols and for the CDS-3M primer but this is not a normalized library. Fifty micrograms of total RNA was extracted from 108 mg of tissue using the Qiagen RNeasy Mini Plant Kit. First strand cDNA was prepared from 1.5 micrograms of total RNA, SuperScript III reverse transcriptase (Invitrogen), CDS-3M primer (Evrogen), and the SMART IV Oligonucleotide (Clontech). Second strand cDNA was prepared by LD-PCR with Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, Finland), digested with SfiI, size fractionated, and cDNA larger than approx. 500 bp was ligated into SfiI-digested pDNR-LIB (Clontech). The ligation mixture was transformed into ElectroMAX DH10B T1 Phage-Resistant electro-competent cells (Invitrogen). Titre of the primary library (1 mL) was approx. 1x10^5 cfu/mL and was not amplified.

text: E. coli DH10B

text: Dendroctonus ponderosae

text: Sfi I

text: Sfi I

text: undetermined

text: whole larvae of mixed instar

text: Dpo01-Larvae-Whole-Untreated-Unnorm (DPO01)

text: Plasmid

text: pDNR-LIB


status: PUBLIC

characteristic: External Id

text: University of British Columbia

characteristic: INSDC center name

characteristic: INSDC first public

text: live

characteristic: INSDC status

text: Generic

characteristic: NCBI submission model

characteristic: NCBI submission package

text: A mountain pine beetle-attacked Lodgepole pine (Pinus contorta) tree was felled on Nov. 28, 2006 along the KayKay Forest Service Road northwest of Prince George, BC, Canada (approx. N 54 02.731' W 123 19.109') and the bolts stored at room temperature with waxed sawn ends to reduce dehydration until use. On Jan. 4, 2007, nine live pupae of undetermined sex were manually removed from the bolt and immediately placed whole in liquid nitrogen. An un-normalized, full-length enriched, directionally cloned library was prepared with the Creator SMART cDNA Library Construction Kit (ClonTech, Mountain View, CA, USA) and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, Moscow, Russia). The Evrogen kit was used for some of the protocols and for the CDS-3M primer but this is not a normalized library. Fifty micrograms of total RNA was extracted from 109 mg of tissue using the Qiagen RNeasy Mini Plant Kit. First strand cDNA was prepared from 1.5 micrograms of total RNA, SuperScript III reverse transcriptase (Invitrogen), CDS-3M primer (Evrogen), and the SMART IV Oligonucleotide (ClonTech). Second strand cDNA was prepared by LD-PCR with Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, Finland), digested with SfiI, size fractionated, and cDNA larger than approx. 500 bp was ligated into SfiI-digested pDNR-LIB (ClonTech). The ligation mixture was transformed into ElectroMAX DH10B T1 Phage-Resistant electro-competent cells (Invitrogen). Titre of the primary library (1 mL) was approx. 4.5x10^5 cfu/mL and was not amplified.

characteristic: description

text: E. coli DH10B

tag: attribute

characteristic: lab-host

text: Dendroctonus ponderosae

characteristic: organism

text: Sfi I

text: Sfi I

text: undetermined

characteristic: sex

text: whole pupae

characteristic: tissue-type

text: Dpo02-Pupae-Whole-Untreated-Unnorm (DPO02)

characteristic: title

text: Plasmid

text: pDNR-LIB

characteristic: vector


text: University of British Columbia

text: live

text: Generic

text: A mountain pine beetle-attacked Lodgepole pine (Pinus contorta) tree was felled on Sept. 21, 2006 along the KayKay Forest Service Road northwest of Prince George, BC, Canada (approx. N 54 02.731' W 123 19.109') and the bolts stored at room temperature with waxed sawn ends to reduce dehydration until use. On Nov. 14, 2006, 46 live pupae of undetermined sex were manually removed from the bolt and immediately placed whole in liquid nitrogen. A normalized, full-length enriched, directionally cloned library was prepared with the Creator SMART cDNA Library Construction Kit (Clontech, Mountain View, CA, USA) and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, Moscow, Russia). Total RNA (832 micrograms) was extracted from frozen tissue (591 mg) using the Qiagen RNeasy Mini Plant Kit. mRNA (29 micrograms) was purified from total RNA (823 micrograms) using the Oligotex mRNA Kit (Qiagen). First strand cDNA was prepared from 1.3 micrograms of mRNA, SuperScript III reverse transcriptase (Invitrogen), CDS-3M primer (Evrogen), and the SMART IV Oligonucleotide (Clontech). Second strand cDNA was prepared by LD-PCR with Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, Finland). This cDNA was then normalized with duplex-specific nuclease and amplified following the TRIMMER-DIRECT protocol, digested with SfiI, size fractionated, and the cDNA larger than approx. 500 bp was ligated into SfiI-digested pDNR-LIB (Clontech). The ligation mixture was transformed into ElectroMAX DH10B T1 Phage-Resistant electro-competent cells (Invitrogen). Titre of the primary library (1 mL) was approx. 2x10^5 cfu/mL and was not amplified.

text: E. coli DH10B

text: Dendroctonus ponderosae

text: Sfi I

text: Sfi I

text: undetermined

text: whole pupae

text: Dpo03-Pupae-Whole-Untreated-Norm (DPO03)

text: Plasmid

text: pDNR-LIB


text: University of British Columbia

text: live

text: Generic

text: A mountain pine beetle-attacked Lodgepole pine (Pinus contorta) tree was felled on Sept. 21, 2006 along the KayKay Forest Service Road northwest of Prince George, BC, Canada (approx. N 54 02.731' W 123 19.109') and the bolts stored at room temperature with waxed sawn ends to reduce dehydration until use. On Apr. 15, 2007, 102 live adults of undetermined sex were manually removed from under the bark of a bolt and immediately placed whole in liquid nitrogen. This corresponds to UNBC collection vials #678-687. The antennae from these beetles were subsequently removed while remaining frozen on dry ice. An un-normalized, full-length enriched, directionally cloned library was prepared with the Creator SMART cDNA Library Construction Kit (Clontech, Mountain View, CA, USA) and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, Moscow, Russia). The Evrogen kit was used for some of the protocols and for the CDS-3M primer but this is not a normalized library. Total RNA (2.6 micrograms) was extracted from the frozen antennae (weight unknown) using the Qiagen RNeasy Mini Plant Kit. First strand cDNA was prepared from 225 nanograms of total RNA, SuperScript III reverse transcriptase (Invitrogen), CDS-3M primer (Evrogen), and the SMART IV Oligonucleotide (Clontech). Second strand cDNA was prepared by LD-PCR with Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, Finland), digested with SfiI, size fractionated, and cDNA larger than approx. 500 bp was ligated into SfiI-digested pDNR-LIB (Clontech). The ligation mixture was transformed into ElectroMAX DH10B T1 Phage-Resistant electro-competent cells (Invitrogen). Titre of the primary library (1 mL) was approx. 1.2x10^5 cfu/mL and was not amplified.

text: E. coli DH10B

text: Dendroctonus ponderosae

text: Sfi I

text: Sfi I

text: undetermined

text: Antennae

text: Dpo04-Adult-Antenna-Untreated-Unnorm (DPO04)

text: Plasmid

text: pDNR-LIB


text: University of British Columbia

text: live

text: Generic

text: A mountain pine beetle-attacked Lodgepole pine (Pinus contorta) tree was felled on Nov. 28, 2006 along the KayKay Forest Service Road northwest of Prince George, BC, Canada (approx. N 54 02.731' W 123 19.109') and the bolts stored at room temperature with waxed sawn ends to reduce dehydration until use. On Jan. 3, 2007, 45 live teneral adults of undetermined sex were manually removed from under the bark of a bolt and immediately placed whole in liquid nitrogen. A normalized, full-length enriched, directionally cloned library was prepared with the Creator SMART cDNA Library Construction Kit (Clontech, Mountain View, CA, USA) and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, Moscow, Russia). Total RNA (583 micrograms) was extracted from frozen tissue (542 mg) using the Qiagen RNeasy Mini Plant Kit. mRNA (704 nanograms, low yield due to spill) was purified from total RNA (450 micrograms) using the Oligotex mRNA Kit (Qiagen). First strand cDNA was prepared from 106 nanograms of mRNA, SuperScript III reverse transcriptase (Invitrogen), CDS-3M primer (Evrogen), and the SMART IV Oligonucleotide (Clontech). Second strand cDNA was prepared by LD-PCR with Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, Finland). This cDNA was then normalized with duplex-specific nuclease and amplified following the TRIMMER-DIRECT protocol, digested with SfiI, size fractionated, and the cDNA larger than approx. 500 bp was ligated into SfiI-digested pDNR-LIB (Clontech). The ligation mixture was transformed into ElectroMAX DH10B T1 Phage-Resistant electro-competent cells (Invitrogen). Titre of the primary library (1 mL) was approx. 4x10^4 cfu/mL and was not amplified.

text: E. coli DH10B

text: Dendroctonus ponderosae

text: Sfi I

text: Sfi I

text: undetermined

text: whole teneral adults

text: Dpo05-TAdult-Whole-Untreated-Norm (DPO05)

text: Plasmid

text: pDNR-LIB


text: University of British Columbia

text: live

text: Generic

text: Mountain pine beetle-attacked Lodgepole pine (Pinus contorta) trees were felled in Fall 2006 along the KayKay Forest Service Road northwest of Prince George, BC, Canada (approx. N 54 02.731' W 123 19.109') and the bolts with waxed sawn ends to reduce dehydration later placed in laboratory emergence cages at room temperature. On Mar. 21-26, 2007, emerging adults were collected from the laboratory emergence cages. On Mar. 27, 2007, juvenile hormone III (racemic, Sigma) diluted in hexane to 2 micrograms/microlitre was applied topically (1 microlitre/beetle) to the abdominal venter of 25 male and 25 female beetles using a 10 microlitre syringe. Beetles separated by sex were incubated for 24 h at room temperature with moistened filter paper in covered glass beakers prior to placing the insects whole in liquid nitrogen. An un-normalized, full-length enriched, directionally cloned library was prepared with the Creator SMART cDNA Library Construction Kit (Clontech, Mountain View, CA, USA) and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, Moscow, Russia). The Evrogen kit was used for some of the protocols and for the CDS-3M primer but this is not a normalized library. Total RNA (474 micrograms) was extracted from frozen tissue (505 mg) using the Qiagen RNeasy Mini Plant Kit. mRNA (3.97 micrograms) was purified from total RNA (440 micrograms) using the Oligotex mRNA Kit (Qiagen). First strand cDNA was prepared from 595 nanograms of mRNA, SuperScript III reverse transcriptase (Invitrogen), CDS-3M primer (Evrogen), and the SMART IV Oligonucleotide (Clontech). Second strand cDNA was prepared by LD-PCR with Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, Finland), digested with SfiI, size fractionated, and cDNA larger than approx. 500 bp was ligated into SfiI-digested pDNR-LIB (Clontech). The ligation mixture was transformed into ElectroMAX DH10B T1 Phage-Resistant electro-competent cells (Invitrogen). Titre of the primary library (1 mL) was approx. 1.7x10^6 cfu/mL and was not amplified.

text: E. coli DH10B

text: Dendroctonus ponderosae

text: Sfi I

text: Sfi I

text: male and female

text: whole adults

text: Dpo06-EAdult-Whole-JH24h-Unnorm (DPO06)

text: Plasmid

text: pDNR-LIB


text: University of British Columbia

text: live

text: Generic

text: Mountain pine beetle-attacked Lodgepole pine (Pinus contorta) trees were felled on Nov. 28, 2006 along the KayKay Forest Service Road northwest of Prince George, BC, Canada (approx. N 54 02.731' W 123 19.109') and the bolts with waxed sawn ends to reduce dehydration later placed in laboratory emergence cages. On Mar. 20, 2007, 60 live emerging adults were collected from the laboratory emergence cages and frozen whole in liquid nitrogen. An un-normalized, full-length enriched, directionally cloned library was prepared with the Creator SMART cDNA Library Construction Kit (Clontech, Mountain View, CA, USA) and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, Moscow, Russia). The Evrogen kit was used for some of the protocols and for the CDS-3M primer but this is not a normalized library. Total RNA (467 micrograms) was extracted from frozen tissue (575 mg) using the Qiagen RNeasy Mini Plant Kit. mRNA (5.8 micrograms) was purified from total RNA (419 micrograms) using the Oligotex mRNA Kit (Qiagen). First strand cDNA was prepared from 600 nanograms of mRNA, SuperScript III reverse transcriptase (Invitrogen), CDS-3M primer (Evrogen), and the SMART IV Oligonucleotide (Clontech). Second strand cDNA was prepared by LD-PCR with Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, Finland), digested with SfiI, size fractionated, and cDNA larger than approx. 500 bp was ligated into SfiI-digested pDNR-LIB (Clontech). The ligation mixture was transformed into ElectroMAX DH10B T1 Phage-Resistant electro-competent cells (Invitrogen). Titre of the primary library (1 mL) was approx. 2.3x10^5 cfu/mL and was not amplified.

text: E. coli DH10B

text: Dendroctonus ponderosae

text: Sfi I

text: Sfi I

text: undetermined

text: whole adults

text: Dpo07-EAdult-Whole-Untreated-Unnorm (DPO07)

text: Plasmid

text: pDNR-LIB


text: University of British Columbia

text: live

text: Generic

text: Mountain pine beetle-attacked Lodgepole pine (Pinus contorta) trees were felled in Fall 2006 along the KayKay Forest Service Road northwest of Prince George, BC, Canada (approx. N 54 02.731' W 123 19.109') and the bolts with waxed sawn ends to reduce dehydration later placed in laboratory emergence cages. On Apr. 5-12, 2007, 50 live mountain pine beetle (Dendroctonus ponderosae Hopkins) emerged adults were collected from the laboratory emergence cages and held on moist filter paper at 4oC. These beetles were then treated Apr. 13-18, 2007 with monoterpenes as follows. Each beetle was put into an individual tube (Fisherbrand disposable culture tubes, Borosilicate glass, 13x100 mm). Tubes with the same sex beetles were placed into a 600 ml glass beaker. As the beetles were able to crawl up the sides of the tubes and potentially escape, each tube was covered with a small piece of laboratory film (Parafilm, Pechiney Plastic Packaging, Menasha, WI), with holes poked in it to allow airflow between the tube and the beaker. A monoterpene ((+)-3-carene, myrcene, (racemic)-alpha-pinene, (-)-beta-pinene, or (-)-verbenone) was pipetted (4 microlitres, neat) into the beaker to allow volatilization but no physical contact with any of the beetles. Each beaker was sealed with Parafilm and the top was covered with foil. They were placed in a fumehood for 24 hours after which time the live beetles were frozen whole in liquid nitrogen. An un-normalized, full-length enriched, directionally cloned library was prepared with the Creator SMART cDNA Library Construction Kit (Clontech, Mountain View, CA, USA) and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, Moscow, Russia). The Evrogen kit was used for some of the protocols and for the CDS-3M primer but this is not a normalized library. Total RNA (339 micrograms) was extracted from frozen tissue (485 mg) using the Qiagen RNeasy Mini Plant Kit. mRNA (1.09 micrograms) was purified from total RNA (304 micrograms) using the Oligotex mRNA Kit (Qiagen). First strand cDNA was prepared from 164 nanograms of mRNA, SuperScript III reverse transcriptase (Invitrogen), CDS-3M primer (Evrogen), and the SMART IV Oligonucleotide (Clontech). Second strand cDNA was prepared by LD-PCR with Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, Finland), digested with SfiI, size fractionated, and cDNA larger than approx. 500 bp was ligated into SfiI-digested pDNR-LIB (Clontech). The ligation mixture was transformed into ElectroMAX DH10B T1 Phage-Resistant electro-competent cells (Invitrogen). Titre of the primary library (1 mL) was approx. 1.3x10^6 cfu/mL and was not amplified.

text: E. coli DH10B

text: Dendroctonus ponderosae

text: Sfi I

text: Sfi I

text: male and female

text: whole adults

text: Dpo08-EAdult-Whole-Monoterp-Unnorm (DPO08)

text: Plasmid

text: pDNR-LIB


text: University of British Columbia

text: live

text: Generic

text: Mountain pine beetle-attacked Lodgepole pine (Pinus contorta) trees were felled in Spring 2008 in Jackman Flats Provincial Park, BC, Canada (approx. N 52 55.958' W 119 22.457') and the bolts with waxed sawn ends to reduce dehydration later placed in laboratory emergence cages. Between Apr. 17 and May 10, 2008, 180 live emerging adults were collected from the laboratory emergence cages and held on moist filter paper at 4oC. Between May 8 and May 22, beetles were immobilized on ice, antennectomized, treated topically on their abdominal venter with 10 micrograms of racemic juvenile hormone in 0.5 microlitres acetone and held in the dark in groups of 10 on moist KimWipes in 60 mL plastic Dixie cups for 1, 5 or 16 hr after which time their anterior midguts and adhering fat body were dissected under water and immediately frozen in liquid nitrogen. At each time point, 60 insects were treated with an approximately equal number of sexes. An un-normalized, full-length enriched, directionally cloned library was prepared with the Creator SMART cDNA Library Construction Kit (Clontech, Mountain View, CA, USA) and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, Moscow, Russia). The Evrogen kit was used for some of the protocols and for the CDS-3M primer but this is not a normalized library. Total RNA (482 micrograms) was extracted from the frozen midgut/fat body tissue of 180 insects (weight undetermined) using the Qiagen RNeasy Mini Plant Kit. mRNA (8.2 micrograms) was purified from total RNA (366 micrograms) using the Oligotex mRNA Kit (Qiagen). First strand cDNA was prepared from 1 microgram of mRNA, SuperScript III reverse transcriptase (Invitrogen), CDS-3M primer (Evrogen), and the SMART IV Oligonucleotide (Clontech). Second strand cDNA was prepared by LD-PCR with Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, Finland), digested with SfiI, size fractionated, and cDNA larger than approx. 500 bp was ligated into SfiI-digested pDNR-LIB (Clontech). The ligation mixture was transformed into ElectroMAX DH10B T1 Phage-Resistant electro-competent cells (Invitrogen). Titre of the primary library (2 mL) was approx. 3.1x10^6 cfu/mL and was not amplified.

text: E. coli DH10B

text: Dendroctonus ponderosae

text: Sfi I

text: Sfi I

text: male and female

text: midgut and adhering fat body

text: Dpo09-EAdult-MGFB-JH-Unnorm (DPO09)

text: Plasmid

text: pDNR-LIB


text: University of British Columbia

text: live

text: Generic

text: Mountain pine beetle-attacked Lodgepole pine (Pinus contorta) trees were felled in Spring 2008 in Jackman Flats Provincial Park, BC, Canada (approx. N 52 55.958' W 119 22.457') and the bolts with waxed sawn ends to reduce dehydration later placed in laboratory emergence cages. Between Apr. 17 and May 10, 2008, 180 live emerging adults were collected from the laboratory emergence cages and held on moist filter paper at 4oC. Between May 8 and May 22, beetles were immobilized on ice, antennectomized, treated topically on their abdominal venter with 10 micrograms of racemic juvenile hormone in 0.5 microlitres acetone and held in the dark in groups of 10 on moist KimWipes in 60 mL plastic Dixie cups for 1, 5 or 16 hr after which time their anterior midguts and adhering fat body were dissected under water and immediately frozen in liquid nitrogen. At each time point, 60 insects were treated with an approximately equal number of sexes. A normalized, full-length enriched, directionally cloned library was prepared with the Creator SMART cDNA Library Construction Kit (Clontech, Mountain View, CA, USA) and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, Moscow, Russia). Total RNA (482 micrograms) was extracted from frozen midgut/fat body tissue of 180 insects (weight undetermined) using the Qiagen RNeasy Mini Plant Kit. mRNA (8.2 micrograms) was purified from total RNA (366 micrograms) using the Oligotex mRNA Kit (Qiagen). First strand cDNA was prepared from 1 microgram of mRNA, SuperScript III reverse transcriptase (Invitrogen), CDS-3M primer (Evrogen), and the SMART IV Oligonucleotide (Clontech). Second strand cDNA was prepared by LD-PCR with Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, Finland). This cDNA was then normalized with duplex-specific nuclease and amplified following the TRIMMER-DIRECT protocol, digested with SfiI, size fractionated, and cDNA larger than approx. 500 bp was ligated into SfiI-digested pDNR-LIB (Clontech). The ligation mixture was transformed into ElectroMAX DH10B T1 Phage-Resistant electro-competent cells (Invitrogen). Titre of the primary library (1 mL) was approx. 1.4x10^5 cfu/mL and was not amplified.

text: E. coli DH10B

text: Dendroctonus ponderosae

text: Sfi I

text: Sfi I

text: male and female

text: midgut and adhering fat body

text: Dpo10-EAdult-MGFB-JH-Norm (DPO10)

text: Plasmid

text: pDNR-LIB


text: University of British Columbia

text: live

text: Generic

text: Un-attacked and mountain pine beetle-attacked Lodgepole pine (Pinus contorta) trees were felled on June 16, 2008 in Jackman Flats Provincial Park, BC, Canada (approx. N 52 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends to reduce dehydration were kept in laboratory emergence cages at room temperature. The un-attacked bolts with waxed sawn ends to reduce dehydration were stored at 4oC until several hours before use. Adult beetles that emerged July 7-14, 2008 were sexed and placed on moist filter paper at 4oC until use. On July 22, females were placed headfirst into 4 mm holes drilled into the bark of un-attacked Lodgepole pine bolts and held in place with mesh. After 24 h of feeding, the mesh was removed, one male was placed into the same hole, and the mesh replaced. The bolt was placed vertically at room temperature and ambient natural light for an additional 40 h of feeding before the beetles were manually extracted from the bolt. The anterior midguts and adhering fat body from a total of 34 beetles were dissected under water and immediately frozen in liquid nitrogen. A normalized, full-length enriched, directionally cloned library was prepared with the Creator SMART cDNA Library Construction Kit (Clontech, Mountain View, CA, USA) and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, Moscow, Russia). Total RNA (106 micrograms) was extracted from frozen midgut/fat body tissue of 34 insects (weight undetermined) using the Qiagen RNeasy Mini Plant Kit. First strand cDNA was prepared from 1 microgram of total RNA, SuperScript III reverse transcriptase (Invitrogen), CDS-3M primer (Evrogen), and the SMART IV Oligonucleotide (Clontech). Second strand cDNA was prepared by LD-PCR with Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, Finland). This cDNA was then normalized with duplex-specific nuclease and amplified following the TRIMMER-DIRECT protocol, digested with SfiI, size fractionated, and cDNA larger than approx. 500 bp was ligated into SfiI-digested pDNR-LIB (Clontech). The ligation mixture was transformed into ElectroMAX DH10B T1 Phage-Resistant electro-competent cells (Invitrogen). Titre of the primary library (1 mL) was approx. 6x10^5 cfu/mL and was not amplified.

text: E. coli DH10B

text: Dendroctonus ponderosae

text: Sfi I

text: Sfi I

text: male and female

text: midgut and adhering fat body

text: Dpo11-EAdult-MGFB-Fed-Norm (DPO11)

text: Plasmid

text: pDNR-LIB


text: University of British Columbia

text: live

text: Generic

text: Un-attacked and mountain pine beetle-attacked Lodgepole pine (Pinus contorta) trees were felled on June 16, 2008 in Jackman Flats Provincial Park, BC, Canada (approx. N 52 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends to reduce dehydration were kept in laboratory emergence cages at room temperature. The un-attacked bolts with waxed sawn ends to reduce dehydration were stored at 4oC until several hours before use. Adult beetles that emerged July 7-14, 2008 were sexed and placed on moist filter paper at 4oC until use. On July 22, females were placed headfirst into 4 mm holes drilled into the bark of un-attacked Lodgepole pine bolts and held in place with mesh. After 24 h of feeding, the mesh was removed, one male was placed into the same hole, and the mesh replaced. The bolt was placed vertically at room temperature and ambient natural light for an additional 40 h of feeding before the beetles were manually extracted from the bolt. The anterior midguts and adhering fat body from a total of 34 beetles were dissected under water and immediately frozen in liquid nitrogen. An un-normalized, full-length enriched, directionally cloned library was prepared with the Creator SMART cDNA Library Construction Kit (Clontech, Mountain View, CA, USA) and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, Moscow, Russia). The Evrogen kit was used for some of the protocols and for the CDS-3M primer but this is not a normalized library. Total RNA (106 micrograms) was extracted from frozen midgut/fat body tissue of 34 insects (weight undetermined) using the Qiagen RNeasy Mini Plant Kit. First strand cDNA was prepared from 1 microgram of total RNA, SuperScript III reverse transcriptase (Invitrogen), CDS-3M primer (Evrogen), and the SMART IV Oligonucleotide (Clontech). Second strand cDNA was prepared by LD-PCR with Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, Finland), digested with SfiI, size fractionated, and cDNA larger than approx. 500 bp was ligated into SfiI-digested pDNR-LIB (Clontech). The ligation mixture was transformed into ElectroMAX DH10B T1 Phage-Resistant electro-competent cells (Invitrogen). Titre of the primary library (1 mL) was approx. 5x10^5 cfu/mL and was not amplified.

text: E. coli DH10B

text: Dendroctonus ponderosae

text: Sfi I

text: Sfi I

text: male and female

text: midgut and adhering fat body

text: Dpo12-EAdult-MGFB-Fed-Unnorm (DPO12)

text: Plasmid

text: pDNR-LIB


text: University of British Columbia

text: live

text: Generic

text: Mountain pine beetle-attacked Lodgepole pine (Pinus contorta) trees were felled in Fall 2006 along the KayKay Forest Service Road northwest of Prince George, BC, Canada (approx. N 54 02.731' W 123 19.109') and the bolts with waxed sawn ends to reduce dehydration later placed in laboratory emergence cages at room temperature. Teneral Adults: On Feb. 6, 2007, 21 live adults of undetermined sex were manually removed from under the bark and immediately placed whole in liquid nitrogen. JH III-treated adults: On Mar. 21-26, 2007, 20 live female emerged adults were collected from the laboratory emergence cages. On Mar. 27, 2007, juvenile hormone III (racemic, Sigma) diluted in hexane to 2 micrograms/microlitre was applied topically (1 microlitre/beetle) to the abdominal venter using a 10 microlitre syringe. Beetles were incubated for 24 h at room temperature with moistened filter paper in covered glass beakers prior to placing the insects whole in liquid nitrogen. Monoterpene fumigated adults: On Apr. 6-12, 2007, 50 emerged adults were collected from the laboratory emergence cages and held on moist filter paper at 4oC. These beetles were then treated Apr. 13-18, 2007 with monoterpenes as follows. Each beetle was put into an individual tube (Fisherbrand disposable culture tubes, Borosilicate glass, 13x100 mm). Tubes with the same sex beetles were placed into a 600 ml glass beaker. As the beetles were able to crawl up the sides of the tubes and potentially escape, each tube was covered with a small piece of laboratory film (Parafilm, Pechiney Plastic Packaging, Menasha, WI), with holes poked in it to allow airflow between the tube and the beaker. A monoterpene ((+)-3-carene, myrcene, (racemic)-alpha-pinene, (-)-beta-pinene, or (-)-verbenone) was pipetted (4 microlitres, neat) into the beaker to allow volatilization but no physical contact with any of the beetles. Each beaker was sealed with Parafilm and the top was covered with foil. They were placed in a fumehood for 24 hours after which time the live beetles were frozen whole in liquid nitrogen. Number of beetles treated [M/F] were as follows: (3)-carene [15/10], myrcene [10/10], alpha-pinene [5/10], beta-pinene [18/19], and verbenone [10/10]. On Oct. 21, 2008, the heads from all the above beetles were removed by razor blade while held frozen on dry ice. A normalized, full-length enriched, directionally cloned library was prepared with the Creator SMART cDNA Library Construction Kit (Clontech, Mountain View, CA, USA) and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, Moscow, Russia). Total RNA (30 micrograms) was extracted from 158 heads (238 mg of tissue) using the Qiagen RNeasy Mini Plant Kit. First strand cDNA was prepared from 1 microgram of total RNA, SuperScript III reverse transcriptase (Invitrogen), CDS-3M primer (Evrogen), and the SMART IV Oligonucleotide (Clontech). Second strand cDNA was prepared by LD-PCR with Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, Finland). This cDNA was then normalized with duplex-specific nuclease and amplified following the TRIMMER-DIRECT protocol, digested with SfiI, size fractionated, and cDNA larger than approx. 500 bp was ligated into SfiI-digested pDNR-LIB (Clontech). The ligation mixture was transformed into ElectroMAX DH10B T1 Phage-Resistant electro-competent cells (Invitrogen). Titre of the primary library (2 mL) was approx. 1x10^6 cfu/mL and was not amplified.

text: E. coli DH10B

text: Dendroctonus ponderosae

text: Sfi I

text: Sfi I

text: male and female

text: heads

text: Dpo13-Adult-Head-Norm (DPO13)

text: Plasmid

text: pDNR-LIB


text: University of British Columbia

text: live

text: Generic

text: Eleven freshly attacked Lodgepole pine (Pinus contorta) trees were identified in late summer 2008 on Crown land just west of Tete Jaune Cache, BC, Canada (approx. N 53 03.588' W 119 36.879'). Six live larvae of mixed instar and of undetermined sex were manually removed from the trees each week between Sept. 19 and Nov. 14, 2008, and immediately frozen in liquid nitrogen. A normalized, full-length enriched, directionally cloned library was prepared with the Creator SMART cDNA Library Construction Kit (Clontech, Mountain View, CA, USA) and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, Moscow, Russia). Total RNA (193 micrograms) was extracted from 54 larvae (291 mg of tissue) using the Qiagen RNeasy Mini Plant Kit. mRNA (1.3 micrograms) was purified from total RNA (110 micrograms) using the Oligotex mRNA Kit (Qiagen). First strand cDNA was prepared from 140 nanograms of mRNA, SuperScript III reverse transcriptase (Invitrogen), CDS-3M primer (Evrogen), and the SMART IV Oligonucleotide (Clontech). Second strand cDNA was prepared by LD-PCR with Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, Finland). This cDNA was then normalized with duplex-specific nuclease and amplified following the TRIMMER-DIRECT protocol, digested with SfiI, size fractionated, and cDNA larger than approx. 500 bp was ligated into SfiI-digested pDNR-LIB (Clontech). The ligation mixture was transformed into ElectroMAX DH10B T1 Phage-Resistant electro-competent cells (Invitrogen). Titre of the primary library (2 mL) was approx. 1.5x10^5 cfu/mL and was not amplified.

text: E. coli DH10B

text: Dendroctonus ponderosae

text: Sfi I

text: Sfi I

text: undetermined

text: whole larvae of mixed instar

text: Dpo14-Larvae-Whole-Coldadapting-Norm (DPO14)

text: Plasmid

text: pDNR-LIB


text: University of Georgia

text: live

text: Generic

characteristic: cell-line

text: Pooled 24 and 48 hr post-infection stages

characteristic: dev-stage

text: Cryptosporidium parvum

text: Eco RV

text: Not I

characteristic: strain

text: Pooled Cryptosporidium parvum KSU-1 enriched 24-48hr post-infection, in-vitro non-normalized nanoscale library

text: Plasmid


text: Zhejiang University

characteristic: INSDC last update

text: live

text: Generic

text: Jinchun No.4

characteristic: cultivar

text: Growing stage

text: E.coli DH5a

text: Cucumis sativus

text: EcoRI

text: Leaf

text: Leaf cDNA library of cucumber (Cucumis sativus L.)

text: Plasmid

text: pGEM-T easy


characteristic: External Id

text: Israel Oceanographic and Limnological Research, National Institute of Oceanography

characteristic: INSDC center name

text: live

text: Generic

text: adult

text: Stylophora pistillata

text: hermaphrodite

text: Stylophora rejection subtraction Library

text: Plasmid


text: Genoscope - Centre National de Sequencage

characteristic: INSDC first public

text: live

characteristic: INSDC status

text: Generic

text: The library was made using the ZAP-cDNA Library Construction Kit (Stratagene) and xylem of 17 years old trees. A mixture of 3 genotypes was used.

text: adult tree

text: Quercus robur

text: EcoRI

text: XhoI

text: xylem

text: Pbluescript SK-


text: live

text: Generic

characteristic: NCBI submission model

characteristic: NCBI submission package

text: The Library was made using the ZAP-cDNA Library Construction Kit (Stratagene) and apical buds harvested on 2 years old seedlings during their development from the dormant bud to the elongating shoot. A mixture of 15 genotypes was used.

text: two year old seedlings

text: Quercus petraea

text: EcoRI

text: XhoI

text: apical bud

text: Pbluescript SK-


text: live

text: Generic

text: The Library was made using the ZAP-cDNA Library Construction Kit (Stratagene) and adult trees roots. A mixture of 10 genotypes was used.

characteristic: description

text: adult tree

text: Quercus robur

text: EcoRI

text: XhoI

text: root

text: Pbluescript SK-


text: live

text: Generic

text: The library was made using the ZAP-cDNA Library Construction Kit (Stratagene) and green leaves of 17 years old trees. A mixture of 3 genotypes was used.

text: adult tree

text: Quercus robur

characteristic: organism

text: EcoRI

text: XhoI

text: leave

text: Pbluescript SK-


text: Bowdoin College

text: live

text: Generic

text: Neural

characteristic: cell type

text: Antepenultimate instar

text: Gryllus bimaculatus

text: Prothoracic ganglia

text: Cricket antepenultimate instar 7-days post dennervation Library


text: Life Science College of Liao Ning Normal University

text: live

text: Generic

text: Artemia sinica

text: whole body

characteristic: tissue-type

text: Artemia sinica whole body EST


text: Georgetown University

text: live

text: Generic

text: Aedes albopictus

characteristic: title


text: Universidade Estadual de Campinas - Instituto de Biologia

text: live

text: Generic

text: Total RNA from workers heads of C. gestroi were reverse transcribed utilizing SuperScript II Reverse Transcriptase (Invitrogen), primer Oligo dT and primer Poly G. The cDNAs contained in 2uL of each RT-product were then amplified by LD-PCR using the same primers. PCR products (5uL) were run on 1% agarose gels and stained with ethidium bromide. The amplification products of over 600 bp were then extracted from the agarose gels and ligated into a pDONR-222 vector (Invitrogen). Ligation was transformed by eletroporation into competent E. coli DH10B-cells and selected for kanamicin resistance. Plasmidial DNA was purified and sequenced using M13 primer (5'-GTAAAACGACGGCCAG-3'). High throughput EST sequencing was performed using dye-terminator chemistry on an ABI 3700 sequencer. The EST sequences were base-called and screened for vector sequences using PHRED and cross-match software. Poly-A repeats and low quality sequences were trimmed and the high quality ones (at least 100 pb with Phred>20) were considered.

text: Workers

text: Coptotermes gestroi

text: Heads

text: Worker heads


text: Northwest Fisheries Science Center

text: live

text: Generic

text: 2-year old fish with cortical alveolus stage follicles

text: ovary

characteristic: organ

text: Oncorhynchus kisutch

text: female

characteristic: sex

text: whole ovarian tissue

text: Subtractive ovarian libraries from fed and fasted coho salmon (14 wk)

text: Plasmid

text: TOPO pCR 2.1

characteristic: vector


text: Florida A & M University

text: live

text: Generic

text: Zinfandel

text: Used Differential Display RT-PCR to isolated transcripts induced specifically upon Xylella fastidiosa infestation.

text: Vitis vinifera

text: Leaf

text: Pierce's Disease induced genes (Zinfandel) Differential Display


text: National Institute of Allergy and Infectious Diseases

text: live

text: Generic

text: Filarial nematode parasite of humans. mRNA was prepared from 1,000,000 microfilariae isolated from blood of infected patients. mRNA was converted to cDNA using the SMART cDNA Library Construction Kit (Clontech). The unamplified library contained 2 x 10^6 pfu/ml with an average insert size of 0.5 kb. Microfilariae were provided by Dr. Thomas Nutman, and the library was constructed by Doran Fink. The library is available from Doran Fink, email doran@nih.gov

text: Microfilariae

characteristic: dev-stage

text: Loa loa

text: hermaphrodite

text: Cameroon

characteristic: strain

text: lambda TriplEx2


text: Central Institute of Medicinal and Aromatic Plants (CIMAP, CSIR)

characteristic: INSDC last update

text: live

text: Generic

text: Sampada

characteristic: cultivar

text: PCR-based suppression subtractive hybridization was carried out using the cDNA populations derived from the poly (A)+ mRNA isolated from the capsule wall of genotype Sampada (tester) and the capsule wall of genotype Sujata (driver) of Papaver somniferum and the subtracted cDNA population was cloned on pGEM-T Easy vector.

text: Capsule

text: Papaver somniferum

text: SSH-derived Papaver somniferum genotype Sampada-specific capsule wall cDNA library

text: Plasmid

text: pGEM-T Easy


characteristic: External Id

characteristic: INSDC center name

text: live

text: Generic

text: Sujata

text: PCR-based suppression subtractive hybridization was carried out using the cDNA populations derived from the poly (A)+ mRNA isolated from the capsule wall of genotype Sujata (tester) and the capsule wall of genotype Sampada (driver) of Papaver somniferum and the subtracted cDNA population was cloned on pGEM-T Easy vector.

text: Capsule

text: Papaver somniferum

text: SSH-derived Papaver somniferum genotype Sujata-specific capsule wall cDNA library

text: Plasmid

text: pGEM-T Easy


text: Northwest Fisheries Science Center

characteristic: INSDC first public

text: live

characteristic: INSDC status

text: Generic

text: ovary

text: Oncorhynchus kisutch

text: female

text: follicle/interstitial cell enriched ovarian tissue

text: Coho salmon subtractive libraries from FSH-treated and untreated ovary, follicle/interstitial cell enriched

text: Plasmid

text: TOPO pCR 2.1


text: Northwest Fisheries Science Center

text: live

text: Generic

characteristic: NCBI submission model

characteristic: NCBI submission package

text: ovary

text: Oncorhynchus kisutch

text: female

text: whole ovarian tissue

text: Coho salmon subtractive libraries from FSH-treated and untreated ovary, whole ovarian tissue

text: Plasmid

text: TOPO pCR 2.1


text: Oregon State University

text: live

text: Generic

text: Stratagene Lambda-ZAP-CMV XR library construction kit

characteristic: description

text: Liver

text: Sebastes melanops

characteristic: organism

text: Sebastes melanops cDNA library

text: phagemid

text: pCMV-Script EX


text: Central Institute of Medicinal and Aromatic Plants

text: live

text: Generic

text: Sampada

text: seedling

text: Papaver somniferum

text: subtracted cDNA library of wounding from P. somniferum


text: Ningbo University of Zhejiang Province

text: live

text: Generic

text: Ink Gland

text: Sepiella maindroni

text: EcoR I

text: Xho I

text: cDNA Library from ink gland of squid (Sepiella maindroni)

text: Plasmid


text: MPI for Molecular Genetics

text: live

text: Generic

text: country: Germany:Hannover ; collected_by: Sabrina Simon

text: larvae

text: Baetis sp. AB-2009

text: whole animal

characteristic: tissue-type


text: MPI for Molecular Genetics

text: live

text: Generic

text: country: Germany:Hannover ; collected_by: Sara Khadjeh

text: adult

text: Ischnura elegans

text: muscle

characteristic: title


text: Xishuangbanna Tropical Botanical Garden

text: live

text: Generic

text: country: China

text: 35-55 (DAF)

text: Jatropha curcas

text: embryo

text: Jatropha curcas embryo 35-55 (DAF)


text: Xishuangbanna Tropical Botanical Garden

text: live

text: Generic

text: country: China

text: 56-70 (DAF)

text: Jatropha curcas

text: embryo

text: Jatropha curcas embryo 56-70 (DAF)


text: Xishuangbanna Tropical Botanical Garden

text: live

text: Generic

text: country: China

text: 71-95 (DAF)

text: Jatropha curcas

text: embryo

text: Jatropha curcas embryo 71-95 (DAF)


text: University of Saskatchewan

text: live

text: Generic

text: mixed

text: Mixture of eight cultivars with varying seed phenotypes: Indian Head, Commando, CDC LeMay, CDC Robin, and breeding lines 1899T-50 and 1788-4 (CDC, Univ. Saskatchewan, Saskatoon, Canada) All developmental stages of seeds and very young fertilized pods were harvested from mature plants, and divided into the following lots: very young fertilized ovaries, young ovules, enlarging seeds, cotyledons of fully filled seed, seed coats of fully filled seeds. cDNA library was made from a mixture of equal amounts of mRNA extracted from each of the above tissues.

text: Mixture of seeds at different development stages plus very young fertilized pods

text: seeds and young fertilized ovaries

characteristic: organ

text: Lens culinaris

text: pBluescript SK+ (Stratagene)


text: University of Delhi South Campus

text: live

text: Generic

text: K-2 & AR-12

text: The subtracted cDNA library was constructed from polyA+ RNA by using PCR-Select cDNA Subtraction Kit (Clontech). The subtracted cDNAs were cloned randomly in pGEM-T Easy vector.

text: Mature

text: Morus indica

text: EcoRI

text: EcoRI

text: hermaphrodite

characteristic: sex

text: Leaf

text: Mulberry dehydration stress subtractive cDNA library

text: Plasmid

text: pGEM-T Easy


text: Lanzhou Veterinary Research Institute

text: live

text: Generic

text: country: China

text: Theileria luwenshuni

text: Theileria luwenshuni cDNA library


text: Third Institute of Oceanography, SOA

text: live

text: Generic

text: Subtracted cDNA library of the polychaete Perineris nuntia exposed to benzo(a)pyrene as compared with the non-exposed control constructed by SSH-PCR. Two cDNA libraries were constructed using the SSH PCR. The forward subtracted library was made using mRNA extracted from the worm exposed to benzo(a)pyrene (about 10ug/L) against the control unexposed worms, whereas the reverse subtracted library was made using the mRNA extracted from the control unexposed worm against the worm exposed to benzo(a)pyrene. The exposure experiments were conducted in glass tanks under laboratory conditions (20+-1oC, salinity 30percent, 12hours dark:12hours light, DO >= 6.0 mg L-1) for 96 hours. Exposed and unexposed sandworms (Perineris nuntia) frozen and stored in liquid nitrogen. Total RNA was extracted using RNAiso Reagent (Takara),and poly(A) mRNA was purified using NucleoTrap mRNA purification kit (BD Biosciences). The Clontech PCR-Select cDNA subtraction kit (BD Biosciences) was then used to enrich for differentially expressed genes. The last two steps of suppressive PCR reactions were completed using BD advanced cDNA PCR Kit (BD Biosciences). The enriched cDNA were purified with DNA Fragment Purification Kit (Takara) and cloned into PMD 18-T vector (Takara), then transferred into DH5a competent cells.

text: Adult worm about two month before turning into heteronereis

text: Perinereis nuntia

text: whole worm


text: University Austral of Chile

text: live

text: Generic

text: normalized cDNA fragments bigger than 500bp.

text: in development

text: Nut

text: Gevuina avellana

text: SAR

text: Cotyledon

text: Gevuina avellana nut in development

text: Phage

text: Lambda ExCell

characteristic: vector


text: Departement de Biochimie, Universite de Montreal

text: live

text: Generic

text: Anophryoides haemophila

text: BCL


text: Instituto Biologia Vegetal y Biotecnologia, Universidad de Talca

text: live

text: Generic

text: ripe fruit

characteristic: dev-stage

text: fruit

text: Fragaria chiloensis

text: SSHA

text: pSC-A-amp/kan


text: live

text: Generic

text: large green fruit

text: fruit

text: Fragaria chiloensis

text: SSHB

text: pSC-A-amp/kan


text: live

text: Generic

text: white fruit

text: fruit

text: Fragaria chiloensis

text: SSHC

text: pSC-A-amp/kan


text: live

text: Generic

text: large green fruit

text: fruit

text: Fragaria chiloensis

text: SSHD

text: pSC-A-amp/kan


text: live

text: Generic

text: ripe fruit

text: fruit

text: Fragaria chiloensis

text: SSHE

text: pSC-A-amp/kan


text: live

text: Generic

text: ripe fruit

text: fruit

text: Fragaria chiloensis

text: SSHF

text: pSC-A-amp/kan


text: National Institute for Genetic Engineering and Biotechnology

text: live

text: Generic

text: Mahouti

text: forty_one_day old seedlings were exposed to high salinity (250 mM NaCl) for 10 days.

text: 6th Leaf

characteristic: lab-host

text: Shoot

text: Triticum aestivum

text: HindIII

text: EcoRI

text: Leaf

text: Triticum aestivum seedlings exposed to long term salinity (250 mM NaCl)

text: Plasmid


text: Brazilian Consortium of Coffee Research and Development / FAPESP

characteristic: INSDC last update

text: live

text: Generic

text: Mundo Novo

text: Flower buds in different developmental stages.

text: Coffea arabica

text: Coffea arabica FB1


text: live

text: Generic

text: Mundo Novo

characteristic: cultivar

text: Leaves treated with araquidonic acid.

text: Coffea arabica

text: Leaves treated with araquidonic acid

text: Coffea arabica AR1


characteristic: External Id

text: live

text: Generic

text: Mundo Novo

text: Young leaves from orthotropic branch.

text: Coffea arabica

text: Young leaves from orthotropic branch

text: Coffea arabica LV5


characteristic: INSDC center name

text: live

text: Generic

text: Mundo Novo

text: Coffea arabica

text: Flower buds in different developmental stages

text: Coffea arabica FB4


characteristic: INSDC first public

text: live

text: Generic

text: Mundo Novo

text: Young leaves from orthotropic branch.

text: Coffea arabica

text: Young leaves from orthotropic branch

text: Coffea arabica LV4


text: live

characteristic: INSDC status

text: Generic

text: Mundo Novo

text: Flower buds, fruits stage 1 and 2.

text: Coffea arabica

text: Flower buds, fruits stage 1 and 2

text: Coffea arabica FR1


text: live

text: Generic

characteristic: NCBI submission model

text: Mundo Novo

text: Mature leaves from plagiotropic branches.

text: Coffea arabica

text: Coffea arabica LV8


text: live

text: Generic

characteristic: NCBI submission package

text: Mundo Novo

text: Coffea arabica

text: Coffea arabica FB2


text: live

text: Generic

text: Mundo Novo

text: Flower buds + pinhead fruits + fruits at different stages.

characteristic: description

text: Coffea arabica

text: Coffea arabica FR2


text: live

text: Generic

text: Catuai

text: Stems infected with Xylella spp.

text: Coffea arabica

characteristic: organism

text: Stems infected with Xylella spp

text: Coffea arabica RX1


text: live

text: Generic

text: Catuai

text: Suspension cells with stressed with aluminum.

text: Coffea arabica

characteristic: tissue-type

text: Coffea arabica RT8


text: live

text: Generic

text: Catuai

text: embryogenic calli.

text: Coffea arabica

text: embryogenic calli

text: Coffea arabica EA1

characteristic: title


text: live

text: Generic

text: Catuai

text: suspension cells with mannose Nacl and KCL.

text: Coffea arabica

text: Coffea arabica CS1


text: live

text: Generic

text: Catuai

text: Suspension cells treated with BTH and brassinoesteroids.

text: Coffea arabica

text: Coffea arabica CB1


text: live

text: Generic

text: Catuai

text: Water deficit stresses field plants (pool of tissues).

text: Coffea arabica

text: Coffea arabica SH2


text: live

text: Generic

text: Primary embryogenic calli. Unknown cultivar.

text: Coffea arabica

text: Primary embryogenic calli

text: Coffea arabica PA1


text: live

text: Generic

text: Roots infected with nematodes. Unknown cultivar.

text: Coffea arabica

text: Roots infected with nematodes

text: Coffea arabica NS1


text: University of Missouri-Columbia

text: live

text: Generic

text: This project was funding by the University of Missouri, Food for the 21st Century. Sample Collection: Slaughterhouse-derived oocytes were collected from either sow or gilt ovaries. Both sow (PMPIIs) and gilt (PMPIIg) oocytes were matured for 42-44 hours to metaphase II (MII) arrest. Gilt oocytes were matured in the defined TCM199 with cysteine, FSH, LH and EGF at the University of Missouri. Sow oocytes were purchased from Art, Inc., Madison, WI (formerly BOMED) and matured in a proprietary medium. Both the cumulus cells and the zona pellucida were removed and MII oocytes were selected with even cytoplasm and an extruded polar body. Pools of 100 were washed and snap frozen for poly-A RNA extraction. Library Construction (PCR Protocol): Libraries were constructed by Scott Korte. The amount of mRNA that was recovered from oocytes was quite limiting and was not sufficient for library production with a standard protocol. Therefore, PCR-based protocol was utilized for producing libraries. Poly-A RNA was isolated by using Dynabeads. The mRNA was reverse transcribed with a NotI-tag-dT18 oligonucleotide and a SMART oligonucleotide (Clontech) modified to contain a SalI site to generate full-length cDNA with a sequence complementary to the SMART oligonucleotide. Sequences within the SMART and dT oligo- nucleotides were used as primers to amplify the cDNAs by PCR with pfu turbo polymerase (Stratagene). The resulting PCR products were purified, digested with NotI and SalI and size fractionated by using a Chroma Spin-400 followed by a Chroma Spin-1000 column (Clontech). Purified cDNA from each PCR reaction was ligated into the pCMV-SPORT1 vector. Preliminary Library Characterization: Randomly chosen clones from each library were analyzed by restriction digestion to determine average insert size (96 clones) and by sequencing (~3 96-well plates) to confirm library quality [e.g. the presence of short polyA+ tails, genomic DNA contamination (must be <1%), ribosomal RNA clones (must be <1%), etc.]. Sequencing and Bioinformatics: If the library met quality standards, then 1000 clones total were sequenced to provide a sequence database representing the predominant clones in each library. The clones were sequenced at the University of Missouri-Columbia DNA Core Facility. Bioinformatics work was performed by GK Springer's bioinformatics group (WG Spollen, JE Ries, A Guillen) in Computer Science and Health Management and Informatics Departments at the University of Missouri-Columbia.

text: Sus scrofa

text: pmpIIg

text: Cloning


text: University of Missouri-Columbia

text: live

text: Generic

text: Sus scrofa

text: pmpIIs

text: Cloning


text: University of Missouri-Columbia

text: live

text: Generic

text: Sus scrofa

text: pueo

text: Cloning


text: University of Missouri-Columbia

text: live

text: Generic

text: Sus scrofa

text: puuo

text: Cloning


text: live

text: Generic

text: Funding: The production of ESTs submitted in this project was funded by USDA Grant 2006-35616-16729 entitled 'Development of Ovine Expressed Sequence Tags for the Study of Female Reproduction' to Jonathan Green (Project Director; University of Missouri); Chris Elsik (Georgetown University); Duane Kiesler (University of Missouri); Randall Prather (University of Missouri); Mike Smith (University of Missouri); Gordon Springer (University of Missouri) and Jerry Taylor (University of Missouri). Genetic Source: Cross-bred ewes for the project were purchased from private producers or were acquired from the research flock at the University of Missouri. Samples collected: The samples consisted of the following: germinal vesicle-stage oocytes; in vivo derived embryos (morula and blastocyst); conceptuses [days 10, 13 and 16]; extraembryonic membranes and embryo from day 20; placenta and fetus from day 35; corpora lutea [days 3, 7, 10, 13 (from pregnant and nonpregnant animals) and pregnant days 16, 20 and 35]; ovarian follicles [day 10 (3 mm) and day 15 (5, 6, & 7 mm); neonatal ovary; oviduct (days 0 and 3); endometrium [days 3, 7, 10, 13 (from pregnant and nonpregnant animals) and pregnant days 16, 20 and 35]. Additional samples were collected from hypothalamus, pituitary, spleen, thymus, skeletal muscle, liver, testes and epididymis. Expanded descriptions of how the tissues were collected can be found at the following URL: http://genome.rnet.missouri.edu/Ovine/Methods/. Library construction (Standard Protocol): Total cellular RNA from each sample was isolated by using STAT-60 reagent (Tel-Test, Friendswood, TX) and poly(A)+ RNA was obtained by two rounds of purification with the Oligotex mRNA isolation kit (Qiagen) according to the manufacturer's instructions. Most of the enzymes and other reagents for double stranded cDNA synthesis were from the pCMV-Script XR cDNA library kit (Stratagene Co; La Jolla, CA). Briefly, 5 ug of poly(A)+ RNA was annealed at 37oC with 10 ug of XhoI-tag-dT18 oligonucleotide (5'-GCTGGTGAGCGGCCGCCTCGAG-tag-T18VN - 3') and reverse transcribed at 37oC with Stratascript (Stratagene) reverse transcriptase in the presence of 5-methyl dCTP. The 'tag' represented a tissue/stage-specific seven base sequence identifier present in the oligonucleotide used to prime first-strand synthesis. The cDNA was converted to a double-stranded form by following the manufacturer's recommendations. The double-stranded cDNAs were ligated to EcoR I adapters and digested with Xho I. The cDNAs were size selected by passage through cDNA size fractionation columns (Invitrogen-Life technologies) and ligated directionally into the EcoR I and Xho I sites of the pCMV-Script vector (Statagene) and transformed into a bacterial host. Library construction (PCR Protocol): The amount of mRNA that could be recovered from oocytes, embryos and the day 10 conceptuses was quite limiting. In these cases, a PCR-based protocol was used for library production. This is the same protocol we have used successfully to produce the oocyte/embryo libraries in our porcine and bovine EST projects (http://genome.rnet.missouri.edu/Bovine/; http://genome.rnet.missouri.edu/Swine/). Poly(A)+ RNA was isolated by using the MicroPoly(A) Pure kit from Ambion (cat. #1918). The mRNA was reverse transcribed with Stratascript (Stratagene) in the presence of a XhoI-tag-dT18 oligonucleotide and a SMART oligonucleotide (Clontech; modified to contain a EcoR1 site) to generate full-length cDNA with a sequence complementary to the SMART oligonucleotide. Sequences within the SMART and dT oligonucleotides were used as primers to amplify the cDNAs by PCR with pfu turbo polymerase (Stratagene). The resulting PCR products were purified, digested with EcoR 1 and Xho 1 and size fractionated. Purified cDNA from each PCR reaction were quantified and ligated into the pCMV-Script vector (Statagene). Preliminary Library Characterization: Randomly chosen clones from each library were analyzed by restriction digestion to determine average insert size and further analyzed by sequencing to confirm library quality and to provide a sequence database representing the predominant clones in each library. Once a library was shown to meet quality expectations, it was sequenced in-depth. Clone Requests: Requests for clones should be made to the Director of the University of Missouri DNA Core facility at: ovine@rnet.missouri.edu. Or to the Project Director at: greenjo@missouri.edu

text: Ovis aries

text: plasmid

text: pCMV-Script


text: live

text: Generic

text: Ovis aries

text: oa

text: plasmid

text: pCMV-Script


text: live

text: Generic

text: Ovis aries

text: ocl

text: plasmid

text: pCMV-Script


text: live

text: Generic

text: Ovis aries

text: ocon

text: plasmid

text: pCMV-Script


text: University of Missouri-Columbia

text: live

text: Generic

text: Ovis aries

text: plasmid

text: pCMV-Script


text: live

text: Generic

text: Ovis aries

text: oemb

text: plasmid

text: pCMV-Script


text: live

text: Generic

text: Ovis aries

text: oend

text: plasmid

text: pCMV-Script


text: live

text: Generic

text: Ovis aries

text: ofet

text: plasmid

text: pCMV-Script


text: live

text: Generic

text: Ovis aries

text: ofol

text: plasmid

text: pCMV-Script


text: live

text: Generic

text: Ovis aries

text: ohyp

text: plasmid

text: pCMV-Script


text: live

text: Generic

text: Ovis aries

text: omix

text: plasmid

text: pCMV-Script


text: live

text: Generic

text: Ovis aries

text: onn

text: plasmid

text: pCMV-Script


text: University of Missouri-Columbia

text: live

text: Generic

text: Ovis aries

text: ooocyte

text: plasmid

text: pCMV-Script


text: live

text: Generic

text: Ovis aries

text: oovi

text: plasmid

text: pCMV-Script


text: live

text: Generic

text: Ovis aries

text: opit

text: plasmid

text: pCMV-Script


text: Nanjing Agricultural University

text: live

text: Generic

text: follicular granulosa cells

characteristic: cell type

text: Capra hircus

text: Female

text: Goat follicular granulosa cells


text: Root Biology Center, South China Agricultural University

text: live

text: Generic

text: root

characteristic: organ

text: Glycine max

text: soybean BX10 supression subtractive hybridization library

text: pEGM-T Easy Vector


text: live

text: Generic

text: Leaves from water deficit stresses plants (Conilon)

text: Coffea canephora

text: Coffea canephora SH1


text: live

text: Generic

text: Embriogenic calli (Conilon)

text: Coffea canephora

text: Embriogenic calli (Conilon)

text: Coffea canephora EC1


text: live

text: Generic

text: Leaves - hidric stress (drought resistent clone - Conilon)

text: Coffea canephora

text: Coffea canephora SH3


text: Brazilian Consortium of Coffee Research and Development / FAPESP

text: live

text: Generic

text: Fruits

text: Coffea racemosa

text: Fruits

text: Coffea racemosa FR4


text: live

text: Generic

text: Fruits

text: Coffea racemosa

text: Fruits

text: Coffea racemosa FV1


text: live

text: Generic

text: Fruits

text: Coffea racemosa

text: Fruits

text: Coffea racemosa FV2


text: live

text: Generic

text: Catuai

text: suspension cells, root and leaves with bion

text: Coffea arabica

text: Coffea arabica BP1


text: live

text: Generic

text: Catuai

text: Well-watered field plants (pool of tissues)

text: Coffea arabica

text: Coffea arabica SS1


text: live

text: Generic

text: Catuai

text: Germinating seeds (whole seeds and zygotic embryos)

text: Coffea arabica

text: Coffea arabica EM1


text: live

text: Generic

text: Catuai

text: Non embryogenic calli

text: Coffea arabica

text: Non embryogenic calli

text: Coffea arabica IC1


text: live

text: Generic

text: Catuai

text: Unknown library

text: Coffea arabica

text: Coffea arabica BP2


text: live

text: Generic

text: Catuai

text: Coffea arabica

text: Coffea arabica SI3


text: live

text: Generic

text: Catuai

text: Embryogenic calli

text: Coffea arabica

text: Embryogenic calli

text: Coffea arabica IA2


text: live

text: Generic

text: Zygotic embryo

text: Coffea arabica

text: Zygotic embryo

text: Coffea arabica EB1


text: live

text: Generic

text: germinating seeds

text: Coffea arabica

text: germinating seeds

text: Coffea arabica SI1


text: live

text: Generic

text: Unknown library

text: Coffea arabica

text: Coffea arabica PL1


text: live

text: Generic

text: Roots

text: Coffea arabica

text: Roots

text: Coffea arabica RT3


text: live

text: Generic

text: Unknown library

text: Coffea arabica

text: Coffea arabica CM1


text: live

text: Generic

text: Unknown library

text: Coffea arabica

text: Coffea arabica LM3


text: live

text: Generic

text: Unknown library

text: Coffea arabica

text: Coffea arabica CD1


text: live

text: Generic

text: Non embryogenic calli + 2,4D

text: Coffea arabica

text: Non embryogenic calli + 2,4D

text: Coffea arabica PC1


text: live

text: Generic

text: Hypocotyls treated with BTH

text: Coffea arabica

text: Hypocotyls treated with BTH

text: Coffea arabica CL2


text: live

text: Generic

text: Plantllets treated with araquidonic acid

text: Coffea arabica

text: Coffea arabica LP1


text: live

text: Generic

text: Unknown library

text: Coffea arabica

text: Coffea arabica FP2


text: live

text: Generic

text: germinating seeds

text: Coffea arabica

text: germinating seeds

text: Coffea arabica SI2


text: live

text: Generic

text: Unknown library

text: Coffea arabica

text: Coffea arabica CL1


text: live

text: Generic

text: Unknown library

text: Coffea arabica

text: Coffea arabica RT7


text: live

text: Generic

text: Roots with BTH

text: Coffea arabica

text: Roots with BTH

text: Coffea arabica RT5


text: live

text: Generic

text: Non embryogenic calli

text: Coffea arabica

text: Non embryogenic calli

text: Coffea arabica CA1


text: live

text: Generic

text: Embryogenic calli

text: Coffea arabica

text: Embryogenic calli

text: Coffea arabica IA1


text: live

text: Generic

text: Mundo Novo

text: Unknown library

text: Coffea arabica

text: Coffea arabica FB3


text: live

text: Generic

text: Mundo Novo

text: Mature leaves from plagiotropic branches

text: Coffea arabica

text: Coffea arabica LV9


text: live

text: Generic

text: Mundo Novo

text: Leaves infected with leaf miner and coffee leaf rust

text: Coffea arabica

text: Coffea arabica RM1


text: National Institute for Genetic Engineering and Biotechnology

text: live

text: Generic

text: forty_five_day old Salicornia persica seedlings were grown in 300 mM NaCl prior to total RNA extraction

text: 8th leaf

text: Shoot

text: Salicornia persica

text: HindIII

text: EcoRI

text: Leaf

text: 300 mM NaCl treated Salicornia persica library

text: Plasmid

characteristic: vector


text: Guangdong Entomological Institute

text: live

text: Generic

text: This is a subtraction library which constructed by suppression subtractive hybridization (SSH) technique.

text: prepupea

characteristic: dev-stage

text: Apis cerana

text: female

characteristic: sex

text: Apis Cerana prepupea 8 hours after vorroa destructor induction

text: plasmid


text: Guangdong Entomological Institute

characteristic: INSDC last update

text: live

text: Generic

text: prepupea

text: Apis mellifera

text: female

text: Apis mellifera prepupea 8 hours after vorroa destructor induction

text: plasmid


text: Universidad Autonoma de Queretaro

text: live

text: Generic

characteristic: cultivar

text: Sequence obtained from interaction between C. chinense BG-3821 and mixed infections with Geminiviruses PHYVV and PepGMW, using SSH.

text: Capsicum chinense

text: Interactions Pepper-Geminiviruses Library


characteristic: External Id

text: University of Delhi South Campus

text: live

text: Generic

text: The cDNA library was constructed from polyA+ RNA by using Universal PCR amplification method. The cDNAs were cloned randomly in pDRIVE vector.

text: Mature

text: Morus indica

text: EcoRI

text: EcoRI

text: hermaphrodite

text: Root

text: Mulberry Root cDNA Library

text: Plasmid

text: pDRIVE


text: Osmania University

characteristic: INSDC center name

text: live

text: Generic

text: Arachis seeds were surface sterilized with Mercuric chloride and coated with Arachis strain specific bradyrhizobium cell suspension. Infected seeds were grown in sterile soil under green house conditions. Total RNA was isolated from the root tissue at nodulation stage employing RNAspin kit from GE Healthcare. Total RNA was employed to reverse transcribe mRNA to cDNA. First and second strand synthesis was performed according to the protocol provided in the SMART cDNA library construction kit from BD Biosciences. The cDNA library was fractionated by agarose gel electrophoresis. The library DNA in the molecular weight range of 0.5Kb to 2.5Kb was gel purified and subsequently employed to ligate to pTZ57R/T vector from InsTAclone PCR Cloning Kit (Fermentas). Ligation mix was used to transform competent XL1-Blue E.coli cells. The positive clones were sequenced employing M13 forward primer (5'-GTAAAACGACGGCCAGT- 3')

text: Nodulation Stage

text: E.coli

characteristic: lab-host

text: Roots

text: Arachis hypogaea

text: Hermaphrodite

text: Kadiri

characteristic: strain

text: whole root tissue

text: Arachis hypogaea L. root cDNA library at nodulation stage

text: Plasmid


text: Universiti Kebangsaan Malaysia, Faculty of Science and Technology

characteristic: INSDC first public

text: live

characteristic: INSDC status

text: Generic

characteristic: NCBI submission model

text: The sequences were obtained from Subtractive Hybridization Library from Salicylic Acid treated leaves (a pool of young and old leaves). The clones were obtained by deep sequencing.

text: pool of young and old

text: Mitragyna speciosa

text: salicylic acid treated leaves

text: Mitragyna speciosa salicylic acid treated leaves

text: pGEMT Easy


text: Central Institute of Medicinal and Aromatic Plants (CIMAP, CSIR)

text: live

text: Generic

characteristic: NCBI submission package

text: Trichome

text: CIM Arogya

text: Poly (A)+ mRNA isolated (at the time of maximal in planta artemisinin content) from the leaf trichomes of a mature Artemisia annua (cv. CIM Arogya) plant growing in the CIMAP farm at Lucknow under the National Gene Bank for Medicinal and Aromatic Plants, was used for preparing the cDNA library in Lambda Zap Express vector (Stratagene)

characteristic: description

text: Mature field-growing plant

text: Leaf

text: Artemisia annua

characteristic: organism

text: Artemisia annua leaf trichome cDNA library in Lambda Zap Express vector

text: Phagemid

text: pBK-CMV


text: live

text: Generic

text: adult worm

text: Sipunculus nudus

text: epidermis, muscle and coelomic fluid

characteristic: tissue-type

text: Sipunculus nudus epidermis, muscle and coelomic fluid adult worm

characteristic: title


text: The University of Tokyo, Asian Natural Environmental Science Center (ANESC)

text: live

text: Generic

text: The RNA isolated from Chloris virgata plants treated with 100 mM NaHCO(3) for 24 h was used to generate this cDNA library.

text: Chloris virgata

text: whole plant

text: cDNA library of Chloris virgata


text: United States Department of Agriculture/Agricultural Research Service

text: live

text: Generic

text: Sedentary parasitic females of reniform nematode were collected from cotton roots and total RNA extracted. Approx. 50 ng of total RNA was reverse-transcribed and amplified using the SMARTTM PCR cDNA Synthesis Kit (Clontech). The amplifed cDNA was used to create the library using the CreatorTM SMARTTM cDNA Library Construction Kit (Clontech).

text: Rotylenchulus reniformis

text: Reniform nematode sedentary parasitic female library

text: pDNR-LIB

characteristic: vector


text: Facultad de Medicina, Universidad de la Republica, UDELAR

text: live

text: Generic

text: 200 ng of total RNA extracted from NEJ with the micro to midi RNA Extraction Kit (Invitrogen) were used as starting material for generating the libraries. Non capped RNAs were dephosphorylated, and the complete mRNAs were decapped by a pyrophosphatase treatment, and later ligated to the GeneRacer RNA oligo (Invitrogen). First strand synthesis was performed with the reverse transcriptase Superscript III (Invitrogen) using GeneRacer oligo-dT primer (5'GCTGTCAACGATACGCTACGTAACGGCATGACAGTG(T)18-3'). Next cDNA amplification by PCR was performed using universal forward (GeneRacer 5'Nested: 5'GGACACTGACATGGACTGAAGGAGTA3') and reverse primers (GenerRacer 3'Nested: 5'CGCTACGTAACGGCATGACAGTG3') provided by the GeneRacer kit. PCR was carried out for 30 cycles (94 oC, 45 sec; 68 oC 45 sec; 72 oC 5 min) using Hot Start Taq DNA polymerase (Fermentas). PCR products were analyzed by electrophoresis, recovered from the gel with QIAquick Gel Extraction kit (QIAGEN), and ligated to pCR4-TA cloning vectors (Invitrogen). Ligation was electroporated into One Shot TOP 10 Electrocompetent E. coli (Invitrogen), plated on LB Amp/X-Gal, recombinant clones from the libraries were randomly picked, grown in Circle Growth medium and stocked at -80 oC in 96-well plates in 30 % glycerol till sequencing.

text: newly excysted juvenile

characteristic: dev-stage

text: Fasciola hepatica

text: Topoisomerase I

text: Topoisomerase I

text: Uruguay

text: Liver fluke newly excysted juvenile

text: Plasmid

text: pCR4-TA cloning vector


text: Insitut National Recherche Agronomique

text: live

text: Generic

text: Fragaria x ananassa

text: Fragaria x ananassa library (Denoyes-Rothan B)


text: live

text: Generic

text: capitola

text: Fragaria x ananassa

text: Fragaria x ananassa cv. capitola library (Denoyes-Rothan B)


text: Insitut National Recherche Agronomique

text: live

text: Generic

text: Fragaria vesca subsp. vesca

text: Fragaria vesca subsp. vesca library (Denoyes-Rothan B)


text: Insitut National Recherche Agronomique

text: live

text: Generic

text: Drymocallis fissa

text: Drymocallis fissa library (Denoyes-Rothan B)


text: Insitut National Recherche Agronomique

text: live

text: Generic

text: Potentilla indica

text: Duchesnea indica library (Denoyes-Rothan B)


text: Insitut National Recherche Agronomique

text: live

text: Generic

text: Rosa lucieae

text: Rosa luciae library (Denoyes-Rothan B)


text: DOE Joint Genome Institute

text: live

text: Generic

text: In Vivo Conditions: infected leaves. The library was made from oligo(dT) primed cDNA and cloned into the pCMV-SPORT6 vector (Invitrogen Corp., Carlsbad, CA). PolyA RNA was primed with an oligo dT primer (5'-GACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'), ligated to a SalI adapter (5'-TCGACCCACGCGTCCG and 5'- CGGACGCGTGGG) and digested with NotI. cDNA was size selected from 2-8kb using 1.1% agarose gel electrophoresis then ligated into NotI and SalI-digested pCMV-SPORT6 vector. The library was constructed and sequencing performedat DOE Joint Genome Institute (Walnut Creek, CA).

text: 4 and 8 day post-inoculation

text: Beta vulgaris/Cercospora beticola mixed EST library

text: SalI

text: NotI

text: sugar beet leaves

text: CCCG Cercospora beticola infected sugar beet leaves


text: Nanjing Forestry University

text: live

text: Generic

text: mycelia in liquid culture

text: 'multigermtubi'

text: Drepanopeziza brunnea f. sp. 'multigermtubi'

text: Marssonina brunnea mycelia in liquid culture


text: Indian Institute of Sugarcane Research, ICAR

text: live

text: Generic

text: BO 91

text: A subtractive cDNA library specific to water-logging stress was prepared from poly (A)+ RNA isolated from leaf of control and water-logging stressed sugarcane plant by using Clonetech PCR-Select cDNA Subtraction kit. Differentially expressed cDNAs were cloned into pGEM-T Easy vector.

text: Mature

text: Saccharum hybrid cultivar BO91

text: Eco RI

text: Eco RI

text: Hermaphrodite

characteristic: sex

text: Leaf

text: Sugarcane water-logging stress induced forward SSH library

text: Plasmid

text: pGEM-T Easy (Promega)


text: Marine Living Resources Research Department, KORDI

characteristic: INSDC last update

text: live

text: Generic

text: Skeletal muscle subtractive cDNA library specific for growth stage, Dark-banded rockfish.

text: Sebastes inermis

text: skeletal muscle

text: Dark-banded rockfish growth stage skeletal muscle subtractive cDNA library


text: DOE Joint Genome Institute

text: live

text: Generic

text: The library was made from oligo(dT)-primed cDNA and cloned into vector pDNR-LIB (Clontech Laboratories, Inc., Mountain View, CA) using the SMART system (BD Biosciences, San Jose, CA). To generate the first strand, polyA RNA was primed with an oligo(dT) primer (5'-ATTCTAGAGGCCGAGGCGGCCGACATG(T)30 VN-3', where V = A, G, or C). The second strand was synthesized by PCR using primer 5'-AAGCAGTGGTATCAACGCAGAGTGGCCATTACGGCCGGG-3' followed by digestion of the SfiI sites. cDNA was size selected then ligated into pDNR-LIB. Library is normalized. Library was created and sequencing done by the DOE Joint Genome Institute (Walnut Creek, CA).

text: Brachypodium distachyon

text: SfiIA

text: SfiIB

text: CCXF Brachypodium distachyon abiotic + biotic stress pool

text: pDNR-LIB


text: DOE Joint Genome Institute

text: live

text: Generic

text: Brachypodium distachyon

text: SfiIA

text: SfiIB

text: CCXG Brachypodium distachyon superpool

text: pDNR-LIB


text: DOE Joint Genome Institute

text: live

text: Generic

text: Brachypodium distachyon

text: SfiIA

text: SfiIB

text: flower

text: CCYO Brachypodium distachyon flower + flower drought

text: pDNR-LIB


text: DOE Joint Genome Institute

text: live

text: Generic

text: Brachypodium distachyon

text: SfiIA

text: SfiIB

text: leaf

text: CCYP Brachypodium distachyon leaf + leaf drought

text: pDNR-LIB


text: Marine Scotland, Marine Laboratory

text: live

text: Generic

text: Subtractive library from IPNV challenged fish. Tester: challenged fish, driver: non-challenged control fish. CLONTECH PCR-Select cDNA Subtraction Kit

text: Parr

text: Salmo salar

text: Head kidney

text: Atlantic salmon 4 weeks post IPNV challenge library

text: Plasmid

text: pDrive (Qiagen)


text: Yunnan Agricultural University

text: live

text: Generic

text: Full-length enriched cDNA library constructed using SMART method. Material is a disease-resistent variety obtained from AVRDC (The World Vegetable Center). Plants are grown for a month until there are four to six euphyllas per plant. They are then inoculated with Phytophthora infestans. After 3-4 days, when signs of disease are seen, RNA is obtained from the leaves. Growth conditions: Temperature: day/night23oC/18oC, Relative humidity: day/night50%/70%, Light application time: 14h/d, 15000Lx .

text: Solanum lycopersicum

text: euphylla

text: Lycopersicon esculentum leaf inoculated with Phytophthora infestans


text: University of Missouri-Columbia

text: live

text: Generic

text: Funding: The production of ESTs submitted in this project was funded by USDA Grant 2006-35616-16729 entitled Development of Ovine Expressed Sequence Tags for the Study of Female Reproduction to Jonathan Green (Project Director; University of Missouri); Chris Elsik (Georgetown University); Duane Kiesler (University of Missouri); Randall Prather (University of Missouri); Mike Smith (University of Missouri); Gordon Springer (University of Missouri) and Jerry Taylor (University of Missouri). Genetic Source: Cross-bred ewes for the project were purchased from private producers or were acquired from the research flock at the University of Missouri. Samples collected: The samples consisted of the following: germinal vesicle-stage oocytes; in vivo derived embryos (morula and blastocyst); conceptuses [days 10, 13 and 16]; extraembryonic membranes and embryo from day 20; placenta and fetus from day 35; corpora lutea [days 3, 7, 10, 13 (from pregnant and nonpregnant animals) and pregnant days 16, 20 and 35]; ovarian follicles [day 10 (3 mm) and day 15 (5, 6, & 7 mm); neonatal ovary; oviduct (days 0 and 3); endometrium [days 3, 7, 10, 13 (from pregnant and nonpregnant animals) and pregnant days 16, 20 and 35]. Additional samples were collected from hypothalamus, pituitary, spleen, thymus, skeletal muscle, liver, testes and epididymis. Expanded descriptions of how the tissues were collected can be found at the following URL: http://genome.rnet.missouri.edu/Ovine/Methods/. Library construction (Standard Protocol): Total cellular RNA from each sample was isolated by using STAT-60 reagent (Tel-Test, Friendswood, TX) and poly(A)+ RNA was obtained by two rounds of purification with the Oligotex mRNA isolation kit (Qiagen) according to the manufacturer's instructions. Most of the enzymes and other reagents for double stranded cDNA synthesis were from the pCMV-Script XR cDNA library kit (Stratagene Co; La Jolla, CA). Briefly, 5 ug of poly(A)+ RNA was annealed at 37oC with 10 ug of XhoI-tag-dT18 oligonucleotide (5'-GCTGGTGAGCGGCCGCCTCGAG-tag-T18VN - 3') and reverse transcribed at 37oC with Stratascript (Stratagene) reverse transcriptase in the presence of 5-methyl dCTP. The 'tag' represented a tissue/stage-specific seven base sequence identifier present in the oligonucleotide used to prime first-strand synthesis. The cDNA was converted to a double-stranded form by following the manufacturer's recommendations. The double-stranded cDNAs were ligated to EcoR I adapters and digested with Xho I. The cDNAs were size selected by passage through cDNA size fractionation columns (Invitrogen-Life technologies) and ligated directionally into the EcoR I and Xho I sites of the pCMV-Script vector (Statagene) and transformed into a bacterial host. Library construction (PCR Protocol): The amount of mRNA that could be recovered from oocytes, embryos and the day 10 conceptuses was quite limiting. In these cases, a PCR-based protocol was used for library production. This is the same protocol we have used successfully to produce the oocyte/embryo libraries in our porcine and bovine EST projects (http://genome.rnet.missouri.edu/Bovine/; http://genome.rnet.missouri.edu/Swine/). Poly(A)+ RNA was isolated by using the MicroPoly(A) Pure kit from Ambion (cat. #1918). The mRNA was reverse transcribed with Stratascript (Stratagene) in the presence of a XhoI-tag-dT18 oligonucleotide and a SMART oligonucleotide (Clontech; modified to contain a EcoR1 site) to generate full-length cDNA with a sequence complementary to the SMART oligonucleotide. Sequences within the SMART and dT oligonucleotides were used as primers to amplify the cDNAs by PCR with pfu turbo polymerase (Stratagene). The resulting PCR products were purified, digested with EcoR 1 and Xho 1 and size fractionated. Purified cDNA from each PCR reaction were quantified and ligated into the pCMV-Script vector (Statagene). Preliminary Library Characterization: Randomly chosen clones from each library were analyzed by restriction digestion to determine average insert size and further analyzed by sequencing to confirm library quality and to provide a sequence database representing the predominant clones in each library. Once a library was shown to meet quality expectations, it was sequenced in-depth. Clone Requests: Requests for clones should be made to the Director of the University of Missouri DNA Core facility at: ovine@rnet.missouri.edu. Or to the Project Director at: greenjo@missouri.edu

text: Ovis aries

text: plasmid

text: pCMV-Script


text: University of Missouri-Columbia

text: live

text: Generic

text: Ovis aries

text: oabicep

text: plasmid

text: pCMV-Script


text: University of Missouri-Columbia

text: live

text: Generic

text: Ovis aries

text: oaliver

text: plasmid

text: pCMV-Script


text: University of Missouri-Columbia

text: live

text: Generic

text: Ovis aries

text: oamepid

text: plasmid

text: pCMV-Script


text: University of Missouri-Columbia

text: live

text: Generic

text: Ovis aries

text: oamtest

text: plasmid

text: pCMV-Script


text: University of Missouri-Columbia

text: live

text: Generic

text: Ovis aries

text: oasplee

text: plasmid

text: pCMV-Script


text: University of Missouri-Columbia

text: live

text: Generic

text: Ovis aries

text: onnova

text: plasmid

text: pCMV-Script


text: University of Missouri-Columbia

text: live

text: Generic

text: Ovis aries

text: onntest

text: plasmid

text: pCMV-Script


text: Huazhong Agricultural University

text: live

text: Generic

text: E. coli BM25.8

text: Prunus mume

text: monoecious

text: Floral organ cDNA library of Mei flower


text: National Research Council

text: live

text: Generic

text: A non-normalized cDNA library was constructed from mRNA of 1 month-old bare root Norway spruce seedlings germinated from a population of open-pollinated seeds. Total RNA for cDNA library construction was prepared following the method of Chang et al, 1993. mRNA was isolated from the total RNA preparation using the PolyATract mRNA Isolation System III (Promega Z5300) according to the manufacturer's instructions, and cDNA was prepared with the Universal RiboClone cDNA Synthesis System (Promega C4360). cDNA products were cloned into pGEM-3Z (Promega P2151) in a random orientation (non-directionally cloning).

text: 1 month old

text: E. coli JM109

text: seedlings

characteristic: organ

text: Picea abies

text: bare root

text: Non-normalized library of 1 month old bare root

text: Plasmid

text: pGEM-3Z (Promega P2151)


characteristic: External Id

text: Facultad de Ciencias Exactas y Naturales, Universidad Nacional de Mar del Plata

text: live

text: Generic

text: Spunta

characteristic: cultivar

text: A directional cDNA library was constructed in the UNIZAP XR cloning vector (ZAP-cDNA Gigapack II Gold Cloning kit, Stratagene) using 3 mg poly(A)+ RNA from 24 h F. eumartii- -infected potato tubers. Differential screening of the cDNA library with a polymerase chain reaction (PCR)-amplified control and subtracted probe was carried out. For preparation of the subtracted probe, 3 mg poly(A)+ RNA from 24 h F. eumartii inoculated tubers were reversed-transcribed using a mixture of oligodT1218 and random hexamers to prime the cDNA synthesis. The poly(A)+ RNA (10 mg) obtained from control tubers (tubers in which PDA sterile disks were kept for 24 h) was photobiotinylated. Next, a subtracted hybridization step between biotinylated poly(A)+ RNA prepared from control tubers and the single-stranded cDNA prepared from fungalinfected tubers was performed. Finally, the subtracted cDNA was converted to double strand molecules, ligated to linkers and amplified by PCR. A control cDNA probe was also prepared following the same procedure, starting from 3 mg poly(A)+ RNA from 24 h PDA sterile diskstreated tubers, and avoiding the subtractive step. Both cDNA probes were labeled with [a-32P]dCTP by random priming, and used directly for the screening of the potato tuber cDNA library. About 500 000 plaques at a density of approximately 50 000 plaques:15 cm plate were screened. Positive recombinant clones were isolated by plaque purification using the same hybridization conditions. In vivo excision was carried out from the selected Uni-ZAP XR cDNA clones, following the manufacturer's instructions (Stratagene) and recombinant pBluescript SK(-) containing colonies were obtained from phagemid infections of XL1-Blue strain (Stratagene).

text: tubers

text: Solanum tuberosum

text: Xho I

text: EcoRI

text: Potato tubers 24 h infected with Fusarium eumartii

text: UNIZAP XR


text: Konkuk University

characteristic: INSDC center name

text: live

text: Generic

text: The brainstem library was constructed by Oligo-capping method. pCNS-D2 vector was used for ligation of double stranded cDNA obtained from the brainstem library. Electro-competent DH10b cells were used for transformation.

text: 5 days old Korean native pig

text: electro-competent DH10b cells

characteristic: lab-host

text: Sus scrofa

text: female

text: Korean native pig

text: brainstem

text: Korean native pig Brainstem by Oligo-capping method


text: Konkuk University

characteristic: INSDC first public

text: live

text: Generic

text: The cerebellum library was constructed by using SMART kit (Clontech, Cat No. 634901). pCNS-D2 vector was used for ligation of double stranded cDNA obtained from the brainstem library. lambdaTriplex 2 vector was used for cloning of double stranded cDNA from cerebellum library. Electro-competent DH10b cells were used for transformation.

text: 5 days old Korean native pig

text: electro-competent DH10b cells

text: Sus scrofa

text: female

text: Korean native pig

characteristic: strain

text: cerebellum

text: Korean native pig Cerebellum by SMART method

text: lTriplex 2


text: Beijing Forestry University

text: live

characteristic: INSDC status

text: Generic

characteristic: NCBI submission model

text: pistil

text: Xanthoceras sorbifolium

text: male

text: pistil

text: Male flowers of Xanthoceras sorbifolium


text: Pusan National University

text: live

text: Generic

characteristic: NCBI submission package

text: breed: thoroughbred

characteristic: description

text: Equus caballus

text: kidney

text: Equus caballus kidney


text: Central Institute of Medicinal and Aromatic Plants (CIMAP, CSIR)

text: live

text: Generic

text: CIM-Arogya

text: Suppression subtractive hybridization was performed between callus tissue (tester) and leaf-trichome (driver) using the Clontech PCR select cDNA subtraction kit as per the manufacturer guidelines. The primary PCR was carried out for 30 cycles and it was followed by a secondary PCR of 15 cycles using Advantage polymerase mixture (Clontech) as per the manufacturer guidelines. Amplicons were cloned into pGEM-T Easy cloning vector and transformed into E.coli competent cells.

text: Callus

text: Artemisia annua

characteristic: organism

text: Artemisia annua leaf-generated callus cDNA library in pGEM-T Easy

text: Plasmid

text: pGEM-T Easy


text: live

text: Generic

text: CIM-Arogya

text: Suppression subtractive hybridization was performed between leaf- trichome (tester) and callus tissue (driver) to identify trichome specific genes using the Clontech PCR select cDNA subtraction kit as per the manufacturer guidelines. The primary PCR was carried out for 30 cycles and it was followed by a secondary PCR of 15 cycles using Advantage polymerase mixture (Clontech) as per the manufacturer guidelines. Amplicons were cloned into pGEM-T Easy cloning vector and transformed into E.coli competent cells.

text: Leaf-trichome

text: Artemisia annua

text: Artemisia annua leaf trichome cDNA library in pGEM-T Easy

characteristic: title

text: Plasmid

text: pGEM-T Easy


text: Agricultural Genetic Engineering Research Institute (AGERI)

text: live

text: Generic

text: country: Egypt

text: Vicia villosa

text: Vicia villosa Egypt library


text: Linkoping University

text: live

text: Generic

text: Physcomitrium patens

text: ESTs from Physcomitrella patens encoding non-specific lipid transfer proteins


text: Linkoping University

text: live

text: Generic

text: Selaginella moellendorffii

text: ESTs from Selaginella moellendorffii encoding non-specific lipid transfer proteins


text: University of Edinburgh

text: live

text: Generic

text: Adult

text: Lumbricus rubellus

text: Hermaphrodite

text: Oleg Georgiev Lumbricus rubellus


text: Swedish Institute for Infectious Disease Control

text: live

text: Generic

text: cDNA library made from Cysticercal stage of Taenia solium (Greene RM, Hancock K, Wilkins PP, Tsang VC. (2000) Taenia solium: molecular cloning and serologic evaluation of 14- and 18-kDa related, diagnostic antigens. J Parasitol. 2000 Oct;86(5):1001-7.)

text: Cysticerca

text: Taenia solium

text: Eco R1

text: Xho 1

text: Taenia solium cysticerca cDNA library

text: Plasmid

text: LamdaZAP

characteristic: vector


text: FNC-CENICAFE, National Coffee Research Center

text: live

text: Generic

text: Beauveria bassiana strain Bb 9024 induced with Hypothenemus hampei. 24h. Sustractiva

text: Micelio creciendo sobre broca

characteristic: dev-stage

text: E.coli

text: Beauveria bassiana

text: Bb 9024

text: Micelio

characteristic: tissue-type

text: Plasmid

text: pGEMT-Easy (Promega)


characteristic: INSDC last update

text: live

text: Generic

text: Beauveria bassiana strain Bb Bb9024 creciendo en broca. Full Length

text: E.coli

text: Beauveria bassiana

text: Bb 9024

text: Micelio

text: Plasmid

text: pDNR-LIB (BD Bioscience)


text: live

text: Generic

text: Beauveria bassiana strain Bb 9119 induced with Hypothenemus hampei. 24h. Sustractiva

text: Micelio creciendo sobre broca

text: E.coli

text: Beauveria bassiana

text: Bb 9119

text: Micelio

text: Plasmid

text: pGEMT-Easy (Promega)


text: live

text: Generic

text: Beauveria bassiana strain Bb 9205 induced with Hypothenemus hampei. 24h. Sustractiva

text: Micelio creciendo sobre broca

text: E.coli

text: Beauveria bassiana

text: Bb 9205

text: Micelio

text: Plasmid

text: pGEMT-Easy (Promega)


text: live

text: Generic

text: Beauveria bassiana strain Bb 9205. Esporas creciendo sobre Hypothenemus hampei por 24 horas. Full

text: Micelio creciendo sobre broca

text: E.coli

text: Beauveria bassiana

text: Bb 9205

text: Micelio

text: Plasmid

text: pDNR-LIB (BD Bioscience)


text: live

text: Generic

text: Beauveria bassiana strain Bb 9205. Micelio de 36 horas creciendo sobre Hypothenemus hampei por 4 hor

text: Micelio creciendo sobre broca

text: E.coli

text: Beauveria bassiana

text: Bb 9205

text: Micelio

text: Plasmid

text: pDNR-LIB (BD Bioscience)


text: USGS

text: live

text: Generic

text: kidney

characteristic: organ

text: Oncorhynchus tshawytscha

text: Chinook salmon kidney PCR_subclone


characteristic: External Id

text: Universidade de Sao Paulo

text: live

text: Generic

text: accessory gland

text: Leptothorax gredleri

text: male

characteristic: sex

text: RDA Leptothorax gredleri

text: Plasmid

text: pGEM


text: DOE Joint Genome Institute

characteristic: INSDC center name

text: live

text: Generic

text: The library was made from oligo(dT) primed cDNA and cloned into the pCMV-SPORT6 vector (Invitrogen Corp., Carlsbad, CA). PolyA RNA was primed with an oligo dT primer (5'-GACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'), ligated to a SalI adapter (5'-TCGACCCACGCGTCCG and 5'- CGGACGCGTGGG) and digested with NotI. cDNA was size selected from 1.2-2kb using 1.1% agarose gel electrophoresis then ligated into NotI and SalI-digested pCMV-SPORT6 vector. The library was constructed and sequencing performed at the DOE Joint Genome Institute (Walnut Creek, CA).

text: 2 days

text: Piromyces sp. E2

text: SalI

text: NotI

text: fungal mat and spores

text: CBSA Piromyces sp. E2 fungal mat and spores


text: DOE Joint Genome Institute

text: live

text: Generic

text: The library was made from oligo(dT) primed cDNA and cloned into the pCMV-SPORT6 vector (Invitrogen Corp., Carlsbad, CA). PolyA RNA was primed with an oligo dT primer (5'-GACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'), ligated to a SalI adapter (5'-TCGACCCACGCGTCCG and 5'- CGGACGCGTGGG) and digested with NotI. cDNA was size selected >2kb using 1.1% agarose gel electrophoresis then ligated into NotI and SalI-digested pCMV-SPORT6 vector. The library was constructed and sequencing performed at the DOE Joint Genome Institute (Walnut Creek, CA).

text: 2 days

characteristic: lab-host

text: Piromyces sp. E2

text: SalI

text: NotI

text: fungal mat and spores

text: CBWA Piromyces sp. E2 fungal mat and spores


text: DOE Joint Genome Institute

characteristic: INSDC first public

text: live

text: Generic

text: The library was made from oligo(dT) primed cDNA and cloned into the pCMV-SPORT6 vector (Invitrogen Corp., Carlsbad, CA). PolyA RNA was primed with an oligo dT primer (5'-GACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'), ligated to a SalI adapter (5'-TCGACCCACGCGTCCG and 5'- CGGACGCGTGGG) and digested with NotI. cDNA was size selected from 0.6-1.2kb using 1.1% agarose gel electrophoresis then ligated into NotI and SalI-digested pCMV-SPORT6 vector. The library was constructed and sequencing performed at the DOE Joint Genome Institute (Walnut Creek, CA).

text: 2 days

text: Piromyces sp. E2

text: SalI

text: NotI

text: fungal mat and spores

text: CBWB Piromyces sp. E2 fungal mat and spores


text: Universite de Caen Basse-Normandie

text: live

text: Generic

text: The library was made from a suppression subtractive hybridization. mRNA extracted from zone A of dogfish testis (germinative zone + spermatogonia stages) were subtracted by mRNA extracted from zone C of dogfish testis (round and young elongating spermatids stages). PCR-select cDNA subtraction kit (Clontech) was used and subtracted cDNA were cloned into pCRII TOPO (Invitrogen). cDNA were sequenced with the universal primers M13 forward or reverse.

text: adult, spermatogonia stages

text: testis

text: Scyliorhinus canicula

text: zone A of testis (germinative zone + spermatogonia stages)

text: Dogfish testis - spermatogonia zone (SSH)

text: plasmid


text: Universite de Caen Basse-Normandie

text: live

text: Generic

text: The library was made from a suppression subtractive hybridization. mRNA extracted from zone C of dogfish testis (round and young elongating spermatids stages) were subtracted by mRNA extracted from zone A of dogfish testis (germinative zone + spermatogonia stages). PCR-select cDNA subtraction kit (Clontech) was used and subtracted cDNA were cloned into pCRII TOPO (Invitrogen). cDNA were sequenced with the universal primers M13 forward or reverse.

text: adult, spermatids stages

text: testis

text: Scyliorhinus canicula

text: male

text: zone C of testis (round and young elongating spermatids stages)

text: Dogfish testis - round spermatids zone (SSH)


text: National Institute of Oceanography

text: live

text: Generic

text: Suppression subtractive hybridization was carried out the products were then cloned into pGEMTeasy vector.

text: 15 day postlarvae

text: Penaeus monodon

text: whole animal

text: Vibrio harveyi challenged Penaeus monodon postlarvae cDNA library

text: pGEMTeasy


text: Central Institute of Medicinal and Aromatic Plants

text: live

text: Generic

text: CIM-Arogya

characteristic: cultivar

text: The cDNA library was constructed from polyA+ RNA by using the ZAP Express cDNA Synthesis and ZAP Express Gigapack III Gold Cloning Kit.

text: Mature

text: Artemisia annua

text: Eco RI

text: Xho I

text: hermaphrodite

text: Trichome

text: Artemisia annua trichome-enriched cDNA library

text: Phagemid

text: pBK-CMV


text: live

characteristic: INSDC status

text: Generic

text: CIM-Arogya

text: The cDNA library was constructed from polyA+ RNA using the Clontech PCR-Select cDNA subtraction kit. A suppression subtractive hybridization was performed between leaf and root plus stem polyA+ RNA. The cDNAs were cloned randomly in pGEM-T Easy vector.

text: Mature

text: Artemisia annua

text: Eco RI

text: Eco RI

text: hermaphrodite

text: Leaf

text: Artemisia annua subtracted cDNA library of leaf tissue

text: Plasmid

text: pGEM-T Easy


text: The institute of crop science research,CAAS

text: live

text: Generic

characteristic: NCBI submission model

text: SSH library was constructed for the root tissue of cultivated soybean genotype 'Wenfeng7'a salt-tolerant cultivar. About one-week-old seedlings when the first pair of leaves expanded fully were used in 1/2hongland solutions containing 0.9% (W/V) NaCl or nothing for control.

text: Glycine max

text: Root

text: Soybean roots SSH cDNA library induced by salt from a salt-tolerant cultivar


text: live

text: Generic

characteristic: NCBI submission package

text: SSH library was constructed for the root tissue of cultivated soybean genotype 'Union' a salt-sensitive cultivar. About one-week-old seedlings when the first pair of leaves expanded fully were used in 1/2hongland solutions containing 0.9% (W/V) NaCl or nothing for control.

text: Glycine max

text: Root

text: soybean roots SSH cDNA library induced by salt from a salt-sensitive cultivar


text: International Academy of Agriculture Science

text: live

text: Generic

text: The plant was cultivated at the field for 3 month. cDNA was inserted into lambda Uni-ZAP XR vector at 5' end with EcoRI and 3' end with XhoI site.

characteristic: description

text: mature

text: E.coli SOLR

text: Agastache rugosa

text: EcoRI

text: XhoI

text: leaf

text: Agastache rugosa leaf lambda phage cDNA library

text: phagemid

text: pBluescript SK(-)


text: Zhejiang Forestry University

text: live

text: Generic

text: Phyllostachys edulis

characteristic: organism

text: Phyllostachys edulis shoots


text: DOE Joint Genome Institute

text: live

text: Generic

text: The library was made from oligo(dT) primed cDNA and cloned into the pCMV-SPORT6 vector (Invitrogen Corp., Carlsbad, CA). PolyA RNA was primed with an oligo dT primer (5'-GACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'), ligated to a SalI adapter (5'-TCGACCCACGCGTCCG and 5'- CGGACGCGTGGG) and digested with NotI. cDNA was size selected from 0.6-2kb using 1.1% agarose gel electrophoresis then ligated into NotI and SalI-digested pCMV-SPORT6 vector. The library was constructed and sequencing performed at the DOE Joint Genome Institute (Walnut Creek, CA).

text: Itaconate production pre-initiation phase

text: Aspergillus terreus ATCC 32359

text: SalI

text: NotI

text: ATCC 32359

characteristic: strain

text: mycelia

text: CFBO Aspergillus terreus pre-itaconate production (L)

characteristic: title


text: DOE Joint Genome Institute

text: live

text: Generic

text: The library was made from oligo(dT) primed cDNA and cloned into the pCMV-SPORT6 vector (Invitrogen Corp., Carlsbad, CA). PolyA RNA was primed with an oligo dT primer (5'-GACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'), ligated to a SalI adapter (5'-TCGACCCACGCGTCCG and 5'- CGGACGCGTGGG) and digested with NotI. cDNA was size selected >2kb using 1.1% agarose gel electrophoresis then ligated into NotI and SalI-digested pCMV-SPORT6 vector. The library was constructed and sequencing performed at the DOE Joint Genome Institute (Walnut Creek, CA).

characteristic: dev-stage

text: Aspergillus terreus ATCC 32359

text: SalI

text: NotI

text: ATCC 32359

text: mycelia

text: CFBP Aspergillus terreus pre-itaconate production (H)

characteristic: vector


text: DOE Joint Genome Institute

characteristic: INSDC last update

text: live

text: Generic

text: Initiation of itaconate production

text: Aspergillus terreus ATCC 32359

text: SalI

text: NotI

text: ATCC 32359

text: mycelia

characteristic: tissue-type

text: CFBS Aspergillus terreus itaconate-initiation (H)


characteristic: External Id

text: DOE Joint Genome Institute

text: live

text: Generic

text: Initiation of itaconate production

text: Aspergillus terreus ATCC 32359

text: SalI

text: NotI

text: ATCC 32359

text: mycelia

text: CFBT Aspergillus terreus itaconate-initiation (L)