text: Oklahoma State University

text: live

text: Generic

text: Bos taurus

text: Embryo

text: bovine embryo cDNA


status: PUBLIC

characteristic: External Id

text: Institute of Chemistry. University of Sao Paulo

characteristic: INSDC center name

characteristic: INSDC first public

characteristic: INSDC last update

text: live

characteristic: INSDC status

text: Generic

characteristic: NCBI submission model

characteristic: NCBI submission package

text: anamorph=Trichoderma reesei; Cloned unidirectionally, 5' end of the cDNA cloned into EcoRI site of pBluescript. Primer: Oligo (dT). Average insert size: 1,2 kb; Uni-ZAP XR Vector system -5' adaptor sequence: 5'GAATTCGGCACGAG3' -3' adaptor sequence: 5'CTCGAGTTTTTTTTTTTTTTTTTT3'

characteristic: description

text: 18 hr Glycerol-grown culture

tag: attribute

characteristic: dev-stage

text: E. coli SOLR cells (kanamycin resistant)

characteristic: lab-host

text: Trichoderma reesei

characteristic: organism

text: EcoRI

text: XhoI

text: Asexual

characteristic: sex

text: QM9414 (ATCC26921)

characteristic: strain

text: Mycelia

characteristic: tissue-type

text: TrEST-A

characteristic: title

text: Phagemid

text: pBluescript SK(+)

characteristic: vector


text: Institution of Laboratory Medicine, Chongqing University of Medical Sciences

text: live

text: Generic

characteristic: cell-line

text: Homo sapiens

text: Human K562 cells


characteristic: External Id

text: live

text: Generic

text: The library was constructed by Claire Murphy, Brandi Chiapelli, and Dr. James McCarter at Washington University, St. Louis. Oligo(dT)-SL1 PCR based library. Ancylostoma caninum SS/L3 cDNA PCR products of size >400 nucleotides containing SL1 on the 5' end and oligo(dT) on the 3' end were non-directionally cloned into pCRII-TOPO(Invitrogen) following the Topo TA cloning protocol. Nematodes were provided by Dr. Prema Arasu (Prema_Arasu@ncsu.edu) of North Carolina State University in Raleigh, NC.

text: Serum stimulated L3

text: Ancylostoma caninum

text: EcoRI

text: EcoRI

text: Ancylostoma caninum L3 SS SL1 TOPO v1 Murphy Chiapelli McCarter

text: plasmid (kanamycin and ampicillin resistant)

text: pCRII-TOPO (Invitrogen)


text: University of Bonn

characteristic: INSDC center name

text: live

text: Generic

text: E. coli XL1-Blue

text: Hebeloma cylindrosporum

text: EcoRI

text: XhoI

text: Mycelia

text: Hebeloma cylindrosporum functional cDNA library

text: pDR 196


text: Jichi Medical School

characteristic: INSDC first public

text: live

text: Generic

text: Cloned by DD-PCR: Primer:[5'TTTTGGCTCC3'], [5'T12MG3']; country=Japan:Tokyo, Arakawa, Higashiogu

text: adult

text: frontal cortex

characteristic: organ

text: Rattus norvegicus

text: male

text: Sprague-Dawley

text: cerebrum

text: JMS frontal dominant cDNA


text: The Institute of Physical and Chemical Research (RIKEN)

characteristic: INSDC last update

text: live

characteristic: INSDC status

text: Generic

text: pooled cell lines ; (cell_line=CRL-1751 WEHI 164), (cell_line=CRL-2116 JC), (cell_line=RCB-0035 WEHI-3), (cell_line=RCB-0464 Meth-A), (cell_line=RCB-0545 OHTA), (cell_line=RCB-0559 K-1 . F1), (cell_line=RCB-1283 B16 melanoma), (cell_type=B cells, cell_line=CRL-1702 WEHI 231), (cell_type=Leydig cells, cell_line=CRL-2065 MLTC-1), (cell_type=Nullipotent stem cell, cell_line=CRL-2070 NE), (tissue_type=bladder, cell_line=RCB-0544 MBT-2), (tissue_type=bone marrow, cell_type=stroma cell, cell_line= CRL-2028 SR-4987), (tissue_type=colon, cell_line=RCB-0549 Cle-H3), (tissue_type=kidney, cell_line=CCL-142 RAG), (tissue_type=submandibular gland, cell_line=CRL-1734 SCA-9 clone 15), (strain=BALB/C, cell_type=B cells, cell_line=CRL-1669 BCL1 Clone 13.20-3B3), (strain=C3H, tissue_type=brain, cell_line=CRL-1443 BC3H1)

text: Mus musculus

text: RIKEN full-length enriched, pooled cell lines


text: live

text: Generic

characteristic: NCBI submission model

characteristic: NCBI submission package

text: CRL-1443 BC3H1

text: Mus musculus

text: brain

text: RIKEN full-length enriched, brain CRL-1443 BC3H1 cDNA


text: live

text: Generic

text: stroma cell

characteristic: cell type

text: CRL-2028 SR-4987

text: 5 months

characteristic: dev-stage

text: Mus musculus

text: female

text: bone marrow

text: RIKEN full-length enriched, bone marrow stroma cell CRL-2028 SR-4987 cDNA


text: live

text: Generic

text: RCB-0464 Meth-A

text: Mus musculus

characteristic: organism

text: female

text: BALB/c

text: sarcoma

text: RIKEN full-length enriched, RCB-0464 Meth-A cDNA


text: live

text: Generic

text: RCB-0545 OHTA

text: embryo

text: Mus musculus

text: ICR

text: RIKEN full-length enriched, RCB-0545 OHTA cDNA


text: live

text: Generic

text: Nullipotent stem cell

text: CRL-2070 NE

text: 7 days embryo

text: Mus musculus

text: RIKEN full-length enriched, Nullipotent stem cell CRL-2070 NE cDNA


text: live

text: Generic

text: CRL-1751 WEHI 164

text: Mus musculus

text: BALB/c

text: RIKEN full-length enriched, CRL-1751 WEHI 164 cDNA


text: live

text: Generic

text: B cells

text: CRL-1669 BCL1 Clone 13.20-3B3

text: Mus musculus

text: BALB/c

text: RIKEN full-length enriched, B cells CRL-1669 BCL1 Clone 13.20-3B3 cDNA


text: live

text: Generic

text: CRL-1734 SCA-9 clone 15

text: Mus musculus

text: male

characteristic: sex

text: Swiss Webster

text: submandibular gland

text: RIKEN full-length enriched, submandibular gland CRL-1734 SCA-9 clone 15 cDNA


text: live

text: Generic

text: RCB-0549 Cle-H3

text: embryo

text: Mus musculus

text: RIKEN full-length enriched, embryo RCB-0549 Cle-H3 cDNA


text: live

text: Generic

text: RCB-0559 K-1.fl

text: spleen

text: Mus musculus

text: DDD

characteristic: strain

text: RIKEN full-length enriched, RCB-0559 K-1 . F1 cDNA


text: live

text: Generic

text: CRL-2116 JC

text: 1.5 years

text: Mus musculus

text: female

text: BALB/cRos

text: mammary gland

characteristic: tissue-type

text: RIKEN full-length enriched, CRL-2116 JC cDNA


text: live

text: Generic

text: CCL-142 RAG

text: kidney

text: Mus musculus

text: BALB/c

text: kidney

text: RIKEN full-length enriched, kidney CCL-142 RAG cDNA

characteristic: title


text: live

text: Generic

text: RCB-0544 MBT-2

text: Mus musculus

text: bladder

text: RIKEN full-length enriched, bladder RCB-0544 MBT-2 cDNA


text: live

text: Generic

text: RCB-1283 B16 melanoma

text: Mus musculus

text: skin

text: RIKEN full-length enriched, RCB-1283 B16 melanoma cDNA


text: The Institute of Physical and Chemical Research (RIKEN)

text: live

text: Generic

text: Leydig cells

text: CRL-2065 MLTC-1

text: Mus musculus

text: testis

text: RIKEN full-length enriched, Leydig cells CRL-2065 MLTC-1 cDNA


text: live

text: Generic

text: RCB-0035 WEHI-3

text: Mus musculus

text: BALB/c

text: blood

text: RIKEN full-length enriched, RCB-0035 WEHI-3 cDNA


text: live

text: Generic

text: B cells

text: CRL-1702 WEHI 231

text: Mus musculus

text: (BALB/c X NZB)F1

text: RIKEN full-length enriched, B cells CRL-1702 WEHI 231 cDNA


text: live

text: Generic

text: 21 days neonate

text: Mus musculus

text: cerebellum

text: RIKEN full-length enriched, 21 days neonate cerebellum


text: live

text: Generic

text: 11 days embryo

text: Mus musculus

text: spinal cord

text: RIKEN full-length enriched, 11 days embryo spinal cord


text: live

text: Generic

text: 13 days embryo

text: Mus musculus

text: brain

text: RIKEN full-length enriched, 13 days embryo brain


text: live

text: Generic

text: adult

text: Mus musculus

text: male

text: gall bladder

text: RIKEN full-length enriched, adult male gall bladder


text: live

text: Generic

text: 13 days embryo

text: Mus musculus

text: spinal cord

text: RIKEN full-length enriched, 13 days embryo spinal cord


text: live

text: Generic

text: 1 month neonate

text: Mus musculus

text: cerebellum

text: RIKEN full-length enriched, 1 month neonate cerebellum


text: live

text: Generic

text: 10 days pregnant adult

text: Mus musculus

text: female

text: ovary and uterus

text: RIKEN full-length enriched, 10 days pregnant adult female ovary and uterus


text: live

text: Generic

text: 16 days neonate

text: Mus musculus

text: male

text: diencephalon

text: RIKEN full-length enriched, 16 days neonate male diencephalon


text: live

text: Generic

text: 16 days neonate

text: Mus musculus

text: male

text: medulla oblongata

text: RIKEN full-length enriched, 16 days neonate male medulla oblongata


text: live

text: Generic

text: adult

text: Mus musculus

text: male

text: vesicular gland

text: RIKEN full-length enriched, adult male vesicular gland


text: live

text: Generic

text: 14 days embryo

text: Mus musculus

text: lung

text: RIKEN full-length enriched, 14 days embryo lung


text: live

text: Generic

text: pooled tissues ; (tissue_type=intestinal mucosa, dev_stage=adult, sex=male), (tissue_type=accessory axillary lymph node, dev_stage=adult, sex=male), (tissue_type=jejunal and colic lymph node, dev_stage=adult, sex=male), (tissue_type=gall bladder, dev_stage=adult, sex=male), (tissue_type=vesicular gland, dev_stage=adult, sex=male), (tissue_type=spinal cord, dev_stage=11 days embryo), (tissue_type=brain, dev_stage=13 days embryo), (tissue_type=spinal cord, dev_stage=13 days embryo), (tissue_type=lung, dev_stage=14 days embryo), (tissue_type=brain, dev_stage=15 days embryo), (tissue_type=ovary and uterus, dev_stage=10 days pregnant adult, sex=female), (tissue_type=cortex, dev_stage=0 day neonate), (tissue_type=cerebellum, dev_stage=1 month neonate), (tissue_type=diencephalon, dev_stage=16 days neonate, sex=male), (tissue_type=medulla oblongata, dev_stage=16 days neonate, sex=male), (tissue_type=cerebellum, dev_stage=21 days neonate), (tissue_type=testis, dev_stage=8 days neonate, sex=male)

characteristic: description

text: Mus musculus

text: RIKEN full-length enriched, pooled tissues, intestinal mucosa, etc.


text: The Institute of Physical and Chemical Research (RIKEN)

text: live

text: Generic

text: adult

text: Mus musculus

text: male

text: accessory axillary lymph node

text: RIKEN full-length enriched, adult male accessory axillary lymph node


text: live

text: Generic

text: 15 days embryo

text: Mus musculus

text: brain

text: RIKEN full-length enriched, 15 days embryo brain


text: live

text: Generic

text: 0 day neonate

text: Mus musculus

text: cortex

text: RIKEN full-length enriched, 0 day neonate cortex


text: live

text: Generic

text: adult

text: Mus musculus

text: male

text: jejunal and colic lymph node

text: RIKEN full-length enriched, adult male jejunal and colic lymph node


text: live

text: Generic

text: adult

text: Mus musculus

text: male

text: intestinal mucosa

text: RIKEN full-length enriched, adult male intestinal mucosa


text: live

text: Generic

text: 8 days neonate

text: Mus musculus

text: male

text: testis

text: RIKEN full-length enriched, 8 days neonate male testis


text: live

text: Generic

text: RCB-0558 LLC

characteristic: cell-line

text: Mus musculus

text: lung

text: RIKEN full-length enriched, lung RCB-0558 LLC cDNA


characteristic: External Id

text: live

text: Generic

text: RCB-0526 Jyg-MC(A)

text: Mus musculus

text: mammary gland

text: RIKEN full-length enriched, mammary gland RCB-0526 Jyg-MC(A) cDNA


characteristic: INSDC center name

text: live

text: Generic

text: RCB-0527 Jyg-MC(B)

text: Mus musculus

text: mammary gland

text: RIKEN full-length enriched, mammary gland RCB-0527 Jyg-MC(B) cDNA


text: Harvard University, Howard Hughes Medical Institute

characteristic: INSDC first public

text: live

text: Generic

text: cDNA made by oligo-dT priming. Size-selected by CHROMA SPIN-1000 (Clontech). Average insert size ~1kb. 5' XhoI site was destroyed after directional cloning. Amplified once. Contact information: Hiroshi Inoue, MD, Metabolism Div. (Alan Permutt Lab), Washington University School of Medicine, Box 8127, 660 South Euclid Ave., St. Louis, MO 63110 E-mail: hinoue@imgate.wustl.edu Tel: 314-362-1916, Fax:314-747-2692.

text: Fetal Pancreas

text: Homo sapiens

text: NotI

text: XhoI

text: Fetal Pancreas (4 Pooled Donors, 18 - 20 weeks, Stratagene #738023)

text: Human Fetal Pancreas 1A

text: pBluescript SK(-)

characteristic: vector


characteristic: INSDC last update

text: live

text: Generic

text: cDNA made by oligo-dT priming. Size-selected on agarose gel. Average insert size ~1kb. 5' XhoI site was destroyed after directional cloning. Amplified once. Contact information: Hiroshi Inoue, MD, Metabolism Div. (Alan Permutt Lab), Washington University School of Medicine, Box 8127, 660 South Euclid Ave., St. Louis, MO 63110 E-mail: hinoue@imgate.wustl.edu Tel: 314-362-1916, Fax:314-747-2692.

text: Fetal Pancreas

text: Homo sapiens

text: NotI

text: XhoI

text: Human Fetal Pancreas 1B

text: pBluescript SK(-)


text: Washington University School of Medicine

text: live

characteristic: INSDC status

text: Generic

text: Williams 82

characteristic: cultivar

text: The cDNA library was constructed from mRNA isolated from very young seeds (less than 20mgs). The library was prepared using the Stratagene pBluescript II SK (+) library construction kit. Complementary DNA was synthesized from mRNA using a primer consisting of a poly(dT) sequence with an Xho I restriction site. Eco RI adaptors were ligated to the blunt-ended cDNA fragments followed by Xho I digestion. The cDNA insert is protected from Xho I digestion via methylation during first strand cDNA sythesis. The cDNA fragments were directionally cloned into the Eco RI-Xho Irestriction site of the pBluescript vector. The ligated cDNA fragments were transformed into E.coli ElectroMax DH10B host cell. The library was constructed by Anu Khanna (Lila Vodkin lab, University of Illinois).

characteristic: lab-host

text: Glycine max

text: EcoRI

text: XhoI

text: young seeds (Williams 82)

text: plasmid

text: pBluescript II SK+


text: live

text: Generic

characteristic: NCBI submission model

characteristic: NCBI submission package

text: 1st strand cDNA was primed with an oligo(dT) primer [ATGTGGCCTTTTTTTTTTTTTTTTT]; double-stranded cDNA was ligated to a DraIII adaptor [TGTTGGCCTACTGG], digested and cloned into distinct DraIII sites of the pME18S-FL3 vector (5' site CACTGTGTG, 3' site CACCATGTG). XhoI should be used to isolate the cDNA insert. Size selection was performed to exclude fragments <1.5kb. Library constructed and donated by Dr. Sumio Sugano (University of Tokyo Institute of Medical Science). Custom primers for sequencing: 5' end primer CTTCTGCTCTAAAAGCTGCG and 3' end primer CGACCTGCAGCTCGAGCACA.

text: adult

text: DH10B (phage resistant)

text: Danio rerio

text: DraIII (CACCATGTG)

text: DraIII (CACTGTGTG)

text: male

text: whole body

text: Sugano SJD adult male

text: phagemid (ampicillin resistant)


text: Scottish Crop Research Institute

text: live

text: Generic

text: Optic

text: Non-normalised library, directionally cloned into pSPORT1. Derived from maternal tissue dissected from developing grains (21 days post anthesis) in glasshouse grown barley plants. Developed as part of the barley transcriptome resources of BBSRC/SEERAD funded cereal IGF (Investigating Gene Function) project.

text: 21 DPA

characteristic: dev-stage

text: Hordeum vulgare subsp. vulgare

characteristic: organism

text: Sal I

text: Not I

text: maternal

text: maternal, 21 DPA, no treatment, cv Optic, EBma07

text: Plasmid


text: Scottish Crop Research Institute

text: live

text: Generic

text: Optic

text: Non-normalised library, directionally cloned into pSPORT1. Derived from shoot and root material of 3 week old etiolated barley plants. N.B. This library has been superseded by EBro07. Developed as part of the barley transcriptome resources of BBSRC/SEERAD funded cereal IGF (Investigating Gene Function) project.

text: 3 week

text: Hordeum vulgare subsp. vulgare

text: Sal I

text: Not I

text: root

text: root, 3 week, reduced light, cv Optic, EBro05

text: Plasmid


text: Scottish Crop Research Institute

text: live

text: Generic

text: Optic

text: Non-normalised library, directionally cloned into pSPORT1. Derived from maternal tissue dissected from developing grains (4 days post anthesis) in glasshouse grown barley plants. Developed as part of the barley transcriptome resources of BBSRC/SEERAD funded cereal IGF (Investigating Gene Function) project.

text: 4 DPA

text: Hordeum vulgare subsp. vulgare

text: Sal I

text: Not I

text: maternal

text: maternal, 4 DPA, no treatment, cv Optic, EBma01

text: Plasmid


text: Scottish Crop Research Institute

text: live

text: Generic

text: Optic

text: Non-normalised library, directionally cloned into pSPORT1. Derived from pistils dissected from developing grains (24 hours post anthesis) in glasshouse grown barley plants. Developed as part of the barley transcriptome resources of BBSRC/SEERAD funded cereal IGF (Investigating Gene Function) project.

text: 1 DPA

text: Hordeum vulgare subsp. vulgare

text: Sal I

text: Not I

text: pistil

text: pistil, 1 DPA, no treatment, cv Optic, EBpi01

text: Plasmid


text: University of Arizona

text: live

text: Generic

text: 12 day old seedlings

text: Zea mays

text: salt stressed roots

text: Zea mays salt stressed roots 12 day old seedlings


text: The Institute of Physical and Chemical Research (RIKEN)

text: live

text: Generic

text: adult

text: Mus musculus

characteristic: strain

text: inner ear

text: RIKEN full-length enriched, adult inner ear


text: US Department of Agriculture (USDA), ARS, PSI

text: live

text: Generic

text: Century

text: cDNA clones from mRNA extracted from soybean cv. Century cotyledons 8 days post germination.

text: Eight days post germination

text: Glycine max

text: Cotyledons

characteristic: tissue-type

text: cDNA Century library

text: phagemid

text: pBluescript SK-


text: live

text: Generic

text: Peking

text: cDNA clones from Glycine max cv. Peking, two days after infection with Soybean Cyst Nematode (SCN) race 3

text: Seedlings

text: Glycine max

text: Roots and leaves

text: cDNA Peking library 2D SCN3

characteristic: title

text: phagemid

text: pBluescript SK-


text: The Institute for Genomic Research (TIGR; www.tigr.org)

text: live

text: Generic

text: Kennebec

text: supplier: Cornell University, Tanksley lab; sequencing; The Institute for Genomic Research. Roots were isolated from in vitro grown stem cuttings on CM medium. Roots were isolated two weeks after placing the stem cuttings from in vitro grown plants on medium.

characteristic: description

text: in vitro grown stem cuttings

text: SOLR

text: Solanum tuberosum

text: EcoRI

text: XhoI

text: roots

text: potato roots

text: pBluescript SK(-)


text: National Institute on Aging/National Institutes of Health

text: live

text: Generic

text: Mouse cDNA project by the Laboratory of Genetics, National Institute on Aging (NIA), Intramural Research Program, NIH (http://lgsun.grc.nia.nih.gov/cDNA). Double-stranded cDNAs were synthesized with an Oligo(dT) primer [Invitrogen: 5'- pGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'] from 48 microgram of total RNA, treated with T4 DNA polymerase, and purified by ethanol-precipitation. The cDNAs were ligated to Lone-linker LL-Sal3 (Ref. Development 127:1737-1749 (2000) [PMID:10725249]), purified by phenol/chloroform, and separated from free linkers by Centricon 100. Then, the cDNAs were digested with SalI and NotI enzymes, and cloned into SalI/NotI site of pSPORT1 plasmid vector. The DH10B E. Coli host was trasnformed with ligation mixture by the standard chemical method. The average insert size is about 1.9 kb. The library was constructed by Yulan Piao (NIA).

text: Newborn

text: brain

characteristic: organ

text: Mus musculus

text: SalI

text: NotI

text: Newborn Brain

text: NIA Mouse Newborn Brain cDNA Library

text: plasmid (ampicillin resistant)

text: pSPORT1 (Invitrogen)


characteristic: External Id

text: live

text: Generic

text: Mouse cDNA project by the Laboratory of Genetics, National Institute on Aging (NIA), Intramural Research Program, NIH (http://lgsun.grc.nia.nih.gov/cDNA). This is a long-transcript enriched cDNA library (Ref. Genome Res. 11:1553-1558 (2001). [PMID:11544199]). In brief,double-stranded cDNAs were synthesized with an Oligo(dT) primer [Invitrogen: 5'- pGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'] from 26 microgram of total RNA, treated with T4 DNA polymerase, and purified by ethanol-precipitation. The cDNAs were ligated to Lone-linker LL-Sal4, purified by phenol/chloroform, and separated from free linkers by Centricon 100. Then, the cDNAs were amplified by long-range high fidelity PCR using Ex Taq polymerase (Takara) with a primer Sal4-S. The products were purified by phenol/chloroform and Centricon 100. The cDNAs were digested with SalI and NotI enzymes, and cloned into SalI/NotI site of pSPORT1 plasmid vector. The DH10B E. coli host was transformed with ligation mixture by the standard chemical method. The average insert size is about 3.0 kb. The library was constructed by Yulan Piao (NIA).

text: Newborn

text: kidney

text: Mus musculus

text: SalI

text: NotI

text: Newborn Kidney

text: NIA Mouse Newborn Kidney cDNA Library (Long)

text: plasmid (ampicillin resistant)

text: pSPORT1 (Invitrogen)


characteristic: INSDC center name

text: live

text: Generic

text: Mouse cDNA project by the Laboratory of Genetics, National Institute on Aging (NIA), Intramural Research Program, NIH (http://lgsun.grc.nia.nih.gov/cDNA). Double-stranded cDNAs were synthesized with an Oligo(dT) primer [Invitrogen: 5'- pGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'] from 24.9 microgram of total RNA, treated with T4 DNA polymerase, and purified by ethanol-precipitation. The cDNAs were ligated to Lone-linker LL-Sal3 (Ref. Development 127:1737-1749 (2000) [PMID: 10725249]), purified by phenol/chloroform, and separated from free linkers by Centricon 100. Then, the cDNAs were digested with SalI and NotI enzymes, and cloned into SalI and NotI site of pSPORT1 plasmid vector. The DH10B E. coli host was transformed with ligation mixture by the chemical method. The average insert size is about 1.8 kb. The library was constructed by Yulan Piao (NIA).

text: Newborn

text: heart

text: Mus musculus

text: SalI

text: NotI

text: Newborn Heart

text: NIA Mouse Newborn Heart cDNA Library

text: plasmid (ampicillin resistant)

text: pSPORT1 (Invitrogen)


text: Washington University School of Medicine

characteristic: INSDC first public

text: live

text: Generic

text: SL1 PCR-based library. Ancylostoma ceylanicum cDNA PCR products of 500-750 nucleotides containing SL1 on the 5' end and oligo(dT) on the 3' end were non-directionally cloned into pCR-XL-TOPO (Invitrogen) following the cloning protocol. The cDNA insert can be excised by digestion with EcoRI. The library was constructed by Dr. Vadim Kapulkin from the University of Colorado at Boulder.

text: adults

text: Ancylostoma ceylanicum

text: EcoRI

text: EcoRI

text: mixed

characteristic: sex

text: Ancylostoma ceylanicum M2 SL1 TOPO Kapulkin

text: Plasmid (kanamycin resistant)

text: pCR-XL-TOPO (Invitrogen)


characteristic: INSDC last update

text: live

text: Generic

text: Toxoplasma RH strain tachyzoites were grown in human foreskin fibroblast cultures in vitro. The library was originally constructed by K.L.Wan, Cambridge University. cDNAs were synthesized from polyA RNAs by oligo d(T) priming and directionally cloned into the EcoRI to XhoI sites of the Lambda ZapII vector using the ZAP-cDNA synthesis kit (Stratagene). The primary cDNA library was mass excised as phagemid using ExAssist helper phage (Stratagene). Phagemid DNA was extracted by phenol-chloroform method, and hybridized against a pool of over-represented ESTs (N>=12, from 5596 previous reads). The subtracted library was electroporated into DH10B (GeneHog, Invitrogen, Inc). WARNING: the library contains a small percentage of cDNAs derived from the human host cells. Library Scource: David Sibley, Washington University.

text: Tachyzoite

text: DH10B (GeneHog, Invitrogen, Inc.)

text: Toxoplasma gondii

text: EcoRI

text: XhoI

text: RH (Type I)

text: TgRH Tachyzoite Subtracted cDNA Library

text: pBluscript SK-

characteristic: vector


text: North Carolina State University

text: live

characteristic: INSDC status

text: Generic

text: The library is from late (summer-August) wood, taken from below the crown of a 20 year old tree. The harvested xylem tissue was on the the cusp between transitional and mature wood. NOTE: The sequences contain a 'cDNA adapter' between the EcoRI site and the start of the EST. The adapter sequence is 'AATTCGGCCATTATGGCC'.

text: late wood

text: Pinus taeda

text: EcoRI

text: Coastal plain loblolly pine from North Carolina

text: primary xylem

text: NXLV (Nsf Xylem Late wood Vertical)

text: pTriplEx


text: US Department of Agriculture, Agriculture Research Service, Pacific West Area, Western Regional Research Center

text: live

text: Generic

characteristic: cultivar

text: Plants were grown in the greenhouse. Spikes were sprayed at anthesis with Fusarium graminearum. Total RNA, and poly(A) RNA were prepared and pooled from infected spike at 0, 6, 12, 24, 36 and 48 hours after inoculation, a cDNA library was made, and the cDNA clones were in vivo excised to give pBluescript phagemids in G. Muehlbauer lab at the University of Minnesota (Kruger, W.M., Muehlbauer, G.J., Pritsch, C., Vance, C.). The cDNA library should contain genes of both wheat and fungal pathogen origin. Plasmid DNA preparations and DNA sequencing were performed in the OD Anderson lab (all other authors).

text: Adult plant

text: E. coli SOLR

characteristic: lab-host

text: Triticum aestivum

text: EcoRI

text: XhoI

text: Spike

text: Wheat Fusarium graminearum infected spike cDNA library

text: Phagemid

text: Lambda Uni-ZAP XR, excised phagemid pBluescript SK


text: Clemson University

text: live

text: Generic

characteristic: NCBI submission model

characteristic: NCBI submission package

text: PI 437654

text: Prepared by suppression subtractive hybridization using root mRNA, 10 hours after soybean cyst nematode's infection as a tester, and the non-infected root mRNA as a driver.

text: Root

text: Glycine max

text: Soybean root forward subtractive cDNA Library

text: plasmid

text: pGEM-T.


text: Clemson University

text: live

text: Generic

text: PI 437654

text: Prepared by suppression subtractive hybridization using the non-infected root mRNA as a tester; the mRNA from soybean cyst nematode infected roots (10 hours after infection) as a driver.

text: Root

text: Glycine max

characteristic: organism

text: Soybean root reverse subtractive cDNA Library

text: plasmid

text: pGEM-T.


text: Telethon Foundation/university of Pisa (Italy)

text: live

text: Generic

text: TOP 10 F'

text: white adipose tissue

text: Mus musculus

text: ECO R1

text: ECO R1

text: male

characteristic: strain

text: White adipose tissue cDNA

text: PCR 2.1 (Invitrogen)


text: Northwestern University

text: live

text: Generic

text: mRNA was isolated from outer hair cells (OHC) and inner hair cells (IHC) of cochlea, and cDNA pools were created by RT-PCR. OHC cDNA was subtracted against IHC cDNA using a PCR-based subtractive technique. The subtracted OHC cDNAs were then cloned into the plasmid pGEM5Z(-) to form an OHC-IHC subtracted library. False positive clones were screened out by differential screening process.

text: mature

characteristic: dev-stage

text: E. coli INV_F'

text: cochlea

text: Meriones unguiculatus

text: NotI

text: EagI

text: male and female

text: Gerbil Outer Hair Cell Subtractive Library


text: The Institute for Genomic Research

text: live

text: Generic

text: At 20-25% parasitemia, blood was collected from BALB/cByJ mice infected with Py17XL parasites, and leukocytes removed by passage over microcrystalline cellulose collumns. Total RNA was isolated using the guanidinium isothiocyanate method, and mRNA isolated using oligo(dT)-cellulose chromatography. First strand cDNA synthesis was completed using a 50-base primer and reverse transcriptase in the presence of 5-methyl dCTP. After second strand synthesis, uneven termini were treated with Pfu DNA polymerase and EcoRI adaptors ligated to the blunt ends. The sample was cleaved with XhoI and separated on a Sephacryl S-500 column. Size-fractionated cDNA was precipitated and ligated to HybriZAP arms directionally using EcoRI-XhoI cleaved arms. After packaging, the phagemid vector (pAD-GAL4) was excised from the HybriZAP vector and plasmid DNA isolated.

text: Asexual blood stages

text: E. coli XL-1 Blue

text: Plasmodium yoelii yoelii

text: PyBS

characteristic: title

text: Yeast activation domain vector


text: National University of Singapore

text: live

text: Generic

text: Poly A+ RNA was isolated from the ovaries of 2 female adult zebrafish of 4-5 month-old and the cDNAs were made using oligo dT primers and inserted into lambda ZAP vector at EcoR1 (5'-cDNA) and XhoI (3'-cDNA) sites using Stratagene's Lambda Uni-ZAP XR cloning system.

characteristic: description

text: 4-5 month-old female adult fish

text: Ovary

text: Danio rerio

text: EcoRI

text: XhoI

text: female

text: Zebrafish Ovary

text: Phagemid

text: Lambda Uni-ZAP XR


text: National University of Singapore

text: live

text: Generic

text: Poly A+ RNA was isolated from the testes of 31 male adult zebrafish of 4-5 month-old and the cDNAs were made using oligo dT primers and inserted into lambda ZAP vector at EcoR1 (5'-cDNA) and XhoI (3'-cDNA) sites using Stratagene's Lambda Uni-ZAP XR cloning system.

text: 4-5 month-old male adult fish

text: Testis

characteristic: organ

text: Danio rerio

text: EcoRI

text: XhoI

text: Male

text: Zebrafish Testis

text: Phagemid

text: Lambda Uni-ZAP XR


characteristic: External Id

text: Institute of Zoology, CAS

text: live

text: Generic

text: Aphis gossypii

text: Aphis gossypii library


text: Washington University School of Medicine

characteristic: INSDC center name

text: live

text: Generic

text: First strand cDNA synthesis was primed using oligo-dT on magnetic beads with an additional primer 5'-gcggccgctaatacgactcacta-taggg-3'. Second strand synthesis was a 3-cycle PCR using the primers 5'-ggccgctaatacgactcactatag-3' and 5'-aagcagtggtaacaacgcagagtacttt-ttttttttttttvn-3'. The cDNA was subsequently amplified in a 7-cycle PCR with the following primers: 5'-ggccgctaatacgactcactatag-3' and 5'-aagcagtggt-aacaacgcag. Deoxy-UMP adaptors were added in a third PCR (5 cycles) and the primers 5'-caucaucaucauggccgctaatacgactcactataggg-3' and 5'-cuacuacuacuaaagcagtggtaacaacgcagagtac-3'. Ends were treated with uracil DNA glycosylase and product with 3' overhangs was annealed to complementary ends of pAMP1. Insert can be excised using EcoRI and NotI. Library constructed by Joe Barnes and Steve Johnson (Washington University).

text: Danio rerio

text: EcoRI

text: NotI

text: male

text: SJD

text: whole body

characteristic: tissue-type

text: Zebrafish SJD adult male II

text: phagemid (ampicillin resistant)


characteristic: INSDC first public

text: live

text: Generic

text: Danio rerio

text: EcoRI

text: NotI

text: SJD

text: pectoral fin

text: SJD adult pectoral fin

text: phagemid (ampicillin resistant)


text: University of California San Diego

characteristic: INSDC last update

text: live

text: Generic

characteristic: cell-line

text: Representational difference analysis of cDNA was used to isolate cDNA sequences induced during the differentiation of CG4 glial progenitor cells into both oligodendrocytes and type-2 astrocytes. cDNA tags were cloned as DpnI fragments into the BamHI site of pBluescript SK+ plasmids.

text: E. Coli

text: Rattus norvegicus

text: Sprague-Dawley

text: Differentiating glial CG-4 cells RDA library

text: Plasmid

text: pBluescript SK+


text: University of South Carolina

text: live

characteristic: INSDC status

text: Generic

text: The mRNA was prepared from antennae of newly eclosed male Aedes. The cDNA-derived blunt-ended PCR products were cloned into the Stratagene pPCR-Script Amp SK(+). Clones were sequenced in both direction using T3 and T7 vector primers.

text: Newly eclosed adults

text: Epicurian Coli XL10-Gold Kan

text: Aedes aegypti

text: SrfI

text: male

characteristic: sex

text: Antenna

text: Aedes aegypti Antennal Library

text: Plasmid

text: pPCR-Script Amp SK(+)

characteristic: vector


text: North Carolina State University

text: live

text: Generic

characteristic: NCBI submission model

text: Giant cells

characteristic: cell type

text: Rutgers

characteristic: cultivar

text: Individual, root-knot induced, tomato giant cells were dissected, and a cDNA bank constructed. This bank was extensively subtracted against healthy root tissue cDNA to generate the current library.

text: Roots

text: Solanum lycopersicum

text: Not I

text: Sfi I

text: Vascular

text: Giant Cell Specific cDNA

text: pGem 11Zf+


text: live

text: Generic

characteristic: NCBI submission package

text: First strand cDNA synthesis was primed using oligo-dT on magnetic beads with an additional primer 5'-gcggccgctaatacgactcacta-taggg-3'. Second strand synthesis was a 3-cycle PCR using the primers 5'-ggccgctaatacgactcactatag-3' and 5'-aagcagtggtaacaacgcagagtacttt-ttttttttttttvn-3'. cDNA was subsequently amplified in a 7-cycle PCR with the following primers: 5'-ggccgctaatacgactcactatag-3' and 5'-aagcagtggt-aacaacgcag. Deoxy-UMP adaptors were added in a third PCR (5 cycles) and the primers 5'-caucaucaucauggccgctaatacgactcactataggg-3' and 5'-cuacuacuacuaaagcagtggtaacaacgcagagtac-3'. Ends were treated with uracil DNA glycosylase and product with 3' overhangs was annealed to complementary ends of pAMP1. Insert can be excised using EcoRI and NotI. Library constructed by Joe Barnes and Steve Johnson (Washington University).

characteristic: lab-host

text: Danio rerio

characteristic: organism

text: EcoRI

text: NotI

text: SJD

characteristic: strain

text: embryo, day 3

text: Zebrafish SJD day 3 embryo

text: phagemid (ampicillin resistant)


text: live

text: Generic

text: First strand cDNA synthesis was primed using oligo-dT on magnetic beads with an additional primer 5'-gcggccgctaatacgactcacta-taggg-3'. Second strand synthesis was a 3-cycle PCR using the primers 5'-ggccgctaatacgactcactatag-3' 5'-aagcagtggtaacaacgcagagtacttt-ttttttttttttvn-3'. cDNA was subsequently amplified in a 7-cycle PCR with the following primers: 5'-ggccgctaatacgactcactatag-3' 5'-aagcagtggt-aacaacgcag. Deoxy-UMP adaptors were added in a third PCR (5 cycles) and the primers 5'-caucaucaucauggccgctaatacgactcactataggg-3' and 5'-cuacuacuacuaaagcagtggtaacaacgcagagtac-3'. Ends were treated with uracil DNA glycosylase and product with 3' overhangs was annealed to complementary ends of pAMP1. Insert can be excised using EcoRI and NotI. Library constructed by Joe Barnes and Steve Johnson (Washington University).

characteristic: description

text: Danio rerio

text: EcoRI

text: NotI

text: embryo, 14 somite

text: Zebrafish C32 14 somite embryo

characteristic: title

text: phagemid (ampicillin resistant)


characteristic: External Id

text: US Army Center for Environmental Health Research / Geo-Centers Inc.

characteristic: INSDC center name

text: live

text: Generic

text: Suppression/subtraction library from livers of male and female medaka treated/untreated with 100 mg/L diethylnitrosamine for 24 h. C clones are untreated. D clones are treated.

text: Adult

characteristic: dev-stage

text: liver

characteristic: organ

text: Oryzias latipes

text: Diethylnitrosamine Treated Medaka

text: pcr 2.1


text: Kazusa DNA Research Institute

characteristic: INSDC first public

characteristic: INSDC last update

text: live

text: Generic

text: isolate=Miyakojima MG-20

text: Lotus japonicus

text: EcoRI

text: XhoI

text: Young plants (two-weeks old)

characteristic: tissue-type

text: Lotus japonicus Young plants (two-weeks old)

text: pBluescriptII SK-


text: Kazusa DNA Research Institute

text: live

characteristic: INSDC status

text: Generic

text: Lotus japonicus

text: EcoRI

text: XhoI

text: ecotype Miyakojima MG-20

text: Pods (20-30 mm in length)

text: Lotus japonicus Pods (20-30 mm in length)

text: pBluescriptII SK-

characteristic: vector


text: Huazhong Agricultural University

text: live

text: Generic

characteristic: NCBI submission model

text: Citrus trifoliata

text: Juvenile early flowering trifoliate orange mutant-subtracted cDNA library


text: National Veterinary Research and Quarantine Servive (NVRQS)

text: live

text: Generic

text: Differentially Expressed gene in Tachyzoite stage of Neospora caninum Korea isolate (Nc-Kr2)

text: Tachyzoites

text: Neospora caninum

text: Korea Isolate Nc-Kr2

text: differentially expressed gene in tachyzoite stage of Neospora caninum Korea isolate Nc-Kr2


text: DOE Joint Genome Institute

text: live

text: Generic

text: In Vivo Conditions: liquid culture. The library was made from oligo(dT)-primed cDNA and cloned into pBIISK-XKAsc (modified pBluescript II SK-; a different MCS has been inserted into the KpnI and NotI sites. The KpnI site has been destroyed.). Library prepared according to Gene (1983)25:263-269 by GATC Biotech; sequencing performed at DOE Joint Genome Institute (Walnut Creek, CA).

text: XL1-Blue MRF'

text: Sporobolomyces roseus IAM 13481

text: EcoRI

text: XhoI

text: IAM 13481

text: CATH Sporobolomyces roseus IAM 13481 whole cells

text: pBSIISK.XKAsc


text: DOE Joint Genome Institute

text: live

text: Generic

text: The library was made from oligo(dT)-primed cDNA and cloned into pDONR222 (Invitrogen Corp., Carlsbad, CA). PolyA RNA was primed with a biotin-attB2-Oligo(dT) primer (5'-GGCGGCCGCACAACTTTGTACAAGAAAGTTGGGT(T)19-3') and the first strand synthesized using Superscript II (Invitrogen Corp., Carlsbad, CA). The second strand was synthesized and an attB1 adaptor ligated to the 5' end (5'-TCGTCGGGGACAACTTTGTACAAAAAAGTTGG-3' and 5'-CCAACTTTTTTGTACAAAGTTGTCCCC-3'). cDNA was size selected from 0.5-1kb using 1% agarose gel electrophoresis and then inserted into the vector using site specific recombination (flanking attB sites on cDNA). Library constructed with RNA harvested from nitrogen limited cells at the Alfred Wegener Institute and sequencing was done at DOE Joint Genome Institute (Walnut Creek, CA).

text: asexual and sexual stages mixed

text: DH10B T1 Phage resistant

text: Aureococcus anophagefferens

text: single cell plant material

text: CATU Aureococcus anophagefferens Single cell plant material, asexual and sexual stages mixed


text: Shimane University

text: live

text: Generic

text: Lentinula edodes

text: Lentinula edodes FMC2


text: Waksman Institute, Rutgers University

text: live

text: Generic

text: cDNAs were reversed transcribed from polyA-selected mRNA from C. brenneri then ligated into BsrG I cut pDONR222 using the Invitrogen Gateway cloning system. The authors gratefully acknowledge the Genome Sequencing Center at The Washington University School of Medicine for providing us with their Caenorhabditis brenneri cDNA library (CPAD1).

text: Mixed stages

text: E. coli DH5alpha

text: Whole animal

text: Caenorhabditis brenneri

text: BsrG I

text: Mixed male and female

characteristic: sex

text: The Washington University Caenorhabditis brenneri cDNA library

text: Plasmid


text: Biotechnology Institute, Ankara University

text: live

text: Generic

characteristic: NCBI submission package

text: Cabernet Sauvignon

characteristic: cultivar

text: Vitis vinifera roots grown in Murashige and Skoog (1962) macro and micronutrients with Gamborg's B5 vitamins (Gamborg et al., 1968) supplemented with 1.5% sucrose, 1% agar under control conditions as well as subjected to cold (0-1.5oC), water-deficit, and 150 mM NaCl stress for 2, 4, 6 h. cDNA Library was constructed using total RNA from cold, drought, 150 mM NaCl stressed and well watered Vitis vinifera roots. Total RNAs from different treatments were extracted and equal amounts were pooled before mRNA selection. Poly(A)+mRNA was isolated from total RNA using the Oligotex Direct mRNA kit (Qiagen). cDNA synthesis was conducted by coverting poly(A)+mRNA to double - stranded cDNA by using primer: 5'-AACTGGAAGAATTCGCGGCCGCTCGCATTTTTTTTTTTTTTTTTTV-3' (V=A,C,G) and Superscript III (Invitrogen). Double stranded cDNAs were size selected (more than 600 bp), adaptored with EcoRI adaptors (aattccgttgctgtcg - Promega # C1291) and digested with NotI. The cDNAs were then directionally cloned into EcoRI-NotI digested pBluescript II SK+ phagemid vector (Stratagene). The total number of white colony forming units (cfu) before amplification was 3x106. Blue colonies (empty vectors) were less than 10%. Purified plasmid DNA from the primary library was converted to single-stranded circles and used as a template for PCR amplification using the T7 and T3 priming sites flanking the cloned cDNA inserts as previously described (Bonaldo M, Lennon G, and Soares MB (1996) Normalization and subtraction: two approaches to facilitate gene discovery. Genome Research 9:791-806). The purified PCR products, representing the entire cloned cDNA population, were used as a driver for normalization. Hybridization between the single-stranded library (50ng) and the PCR products (500ng) was carried out for 44 hours at 30oC. Unhybridized single-stranded DNA circles were separated from hybridized DNA rendered partially double-stranded and electroporated into DH10B cells to generate the normalized library. The total number of clones with insert was: 1.6x106 cfu. Background of empty clones was less than 10%.

text: 10 cm high plants grown in Magenta boxes

text: Vitis vinifera

text: EcoRI

text: XhoI

text: roots

text: VVM

text: Phagemid

text: Lambda Uni-Zap XR, Bluescript SK-


text: Centro de Pesquisas do Cacau - CEPEC - Cx. postal 07

text: live

text: Generic

text: Total RNA was extracted from Theobroma cacao L. pods (TSH1188) inoculated by with M. perniciosa. Inoculated pods were harvested at 24h, 48h, 72h and then every 5 days to 120 days after inoculation. The cDNA library was constructed from pooled total RNA, using the DB SMART Creator cDNA library kit (Clontech), and the cDNA were cloned directionally into the pDNR-LIB plasmid.

text: Tissues were collected at 24h, 48h, 72h, 5 days after inoculation (DAI), 15 DAI, 30 DAI, 45 DAI, 60 DAI, 90 DAI and 120 DAI

text: Pods

text: Theobroma cacao

characteristic: organism

text: Theobroma cacao-Moniliophthora perniciosa pods incompatible interaction library

text: Plasmid

text: pDNR-LIB


text: University of Victoria

text: live

text: Generic

text: Tissue contributors: Carlo Biagi, Mitch Uh and Robert Devlin (DFO, Vancouver, B.C.), Simon Jones (PBS, Nanaimo, B.C.), Seaspring Hatchery (Crofton, B.C.), Rachel Roper (University of Victoria)

text: Salmo salar

text: gill

text: pBlueScriptIISK+


text: University of Victoria

text: live

text: Generic

text: Salmo salar

text: brain

text: pBlueScriptIISK+


text: University of Victoria

text: live

text: Generic

text: Salmo salar

text: skeletal muscle

text: pBlueScriptIISK+


text: University of Victoria

text: live

text: Generic

text: Salmo salar

text: liver

text: pBlueScriptIISK+


text: University of Victoria

text: live

text: Generic

text: Salmo salar

text: whole

text: pBlueScriptIISK+


text: University of Victoria

text: live

text: Generic

text: Salmo salar

text: spleen, kidney, brain


text: University of Victoria

text: live

text: Generic

text: Salmo salar

text: retina

text: pBlueScriptIISK+


text: University of Victoria

text: live

text: Generic

text: Salmo salar

text: cardiac muscle

text: pBlueScriptIISK+


text: University of Victoria

text: live

text: Generic

text: Salmo salar

text: testis

text: pBlueScriptIISK+


text: University of Victoria

text: live

text: Generic

text: Salmo salar

text: gut

text: pBlueScriptIISK+


text: University of Victoria

text: live

text: Generic

text: Salmo salar

text: pyloric caeca

text: pBlueScriptIISK+


text: University of Victoria

text: live

text: Generic

text: Salmo salar

text: esophagus

text: pBlueScriptIISK+


text: University of Victoria

text: live

text: Generic

text: Salmo salar

text: head kidney

text: pBlueScriptIISK+


text: University of Victoria

text: live

text: Generic

text: Tissue contributors: Robert Devlin (DFO, Vancouver, B.C.)

text: Salmo salar

text: pyloric caeca

text: pBlueScriptIISK+


text: University of Victoria

text: live

text: Generic

text: Salmo salar

text: ovary

text: pBlueScriptIISK+


text: University of Victoria

text: live

text: Generic

text: Salmo salar

text: pituitary

text: pBlueScriptIISK+


text: University of Victoria

text: live

text: Generic

text: Salmo salar

text: brain, kidney, spleen

text: pDNR-LIB


text: University of Victoria

text: live

text: Generic

text: Salmo salar

text: spleen

text: pBlueScriptIISK+


text: NRC Institute for Marine Biosciences Industrial Partnership Facility

text: live

text: Generic

text: Gadus morhua

text: pyloric caeca

text: Gadus morhua pyloric caeca library (injected with polyriboinosinic polyribocytidylic acid)


text: DOE Joint Genome Institute

text: live

text: Generic

text: The library was made from oligo(dT)-primed cDNA and cloned into pBluescript SK- (Stratagene, La Jolla, CA). Library constructed and sequencing performed at DOE Joint Genome Institute (Walnut Creek, CA).

text: Chlamydomonas reinhardtii

characteristic: strain

text: CACW Chlamydomonas reinhardtii Strain S1D2 normal


text: University of Washington

text: live

text: Generic

text: Poly A RNA was prepared from total RNA prep. Full-size, double-stranded cDNA was prepared by SMART synthesis and normalized using kamchatka crab duplex-specific nuclease. SfiI-digested cDNA was cloned in SfiIA and SfiIB sites in pAL17.3 plasmid vector. Sequencing was carried out on a template consisting of vector DNA from cDNA-carrying clones selected by blue/white screeing.

characteristic: description

text: placenta

text: Papio anubis

text: female

text: plasmid


text: University of Manchester

text: live

text: Generic

text: Adult

text: Kidney

text: Platichthys flesus

text: Wild type

text: Flounder kidney library

characteristic: title

text: plasmid

text: pTA-easy or pDNR-LIB


characteristic: External Id

text: Shanghai Institute of Planned Parenthood Research

text: live

text: Generic

text: oocyte

characteristic: cell type

text: IV

text: Bufo gargarizans

text: pMyr


text: Forestry and Forest Products Research Institute (FFPRI)

characteristic: INSDC center name

text: live

text: Generic

text: synonym: Populus nigra var. italica

text: mixture of juvenile, mature

characteristic: dev-stage

text: Populus nigra

text: female

text: mixture of leaf, bud, stem, root

text: full-length enriched poplar cDNA library, mixture of leaf, bud, stem, root


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 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 for 1.2-2.5kb using 1% agarose gel electrophoresis, then ligated into pDNR-LIB. Library was created and sequencing performed at DOE Joint Genome Institute (Walnut Creek, CA).

text: embryo < 24 hours

text: Nematostella vectensis

text: SfiI A

text: SfiI B

text: CAAB Nematostella vectensis nemve_P1 Early (embryo < 24 hours) polyA

text: pDNR-LIB


text: DOE Joint Genome Institute

characteristic: INSDC last update

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 from 0.6-1.5kb using 1% agarose gel electrophoresis, then ligated into pDNR-LIB. Library was created and sequencing performed at DOE Joint Genome Institute (Walnut Creek, CA).

text: embryo < 24 hours

text: Nematostella vectensis

text: SfiI A

text: SfiI B

text: CAAD Nematostella vectensis nemve_P1 Early embryo < 24 hours

text: pDNR-LIB


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 from 0.5-1kb using 1% agarose gel electrophoresis, then ligated into pDNR-LIB. Library was created and sequencing performed at DOE Joint Genome Institute (Walnut Creek, CA).

text: embryo late < 24hrs

text: Nematostella vectensis

text: SfiI A

text: SfiI B

text: CAAE Nematostella vectensis nemve_P2 Late < 24 hrs 0.5-1kb

text: pDNR-LIB


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 from 1.5-4kb using 1% agarose gel electrophoresis, then ligated into pDNR-LIB. Library was created and sequencing performed at DOE Joint Genome Institute (Walnut Creek, CA).

text: embryo late < 24hrs

text: Nematostella vectensis

text: SfiI A

text: SfiI B

text: CAAF Nematostella vectensis nemve_P2 Late < 24 hrs 1.5-4kb

text: pDNR-LIB


text: DOE Joint Genome Institute

text: live

text: Generic

text: The library was made from oligo(dT)-primed cDNA and cloned into the pSPORT1 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 1.2-2.5kb using 1.1% agarose gel electrophoresis, then ligated into NotI and SalI-digested pSPORT1 vector. Library was created and sequencing done by the DOE Joint Genome Institute (Walnut Creek, CA).

text: early polyp

text: Nematostella vectensis

text: SalI

text: NotI

text: CACG Nematostella vectensis nemve mixed stage egg - early polyp M


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 0.6-1.5kb using 1.1% agarose gel electrophoresis then ligated into NotI and SalI-digested pCMV-SPORT6 vector. The library was constructed at DOE Joint Genome Institute (Walnut Creek, CA).

text: Early embryo

text: Nematostella vectensis

text: SalI

text: NotI

text: CAGC Nematostella vectensis Nemve Early embryo <24hrs


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 1.2-2kb using 1.1% agarose gel electrophoresis then ligated into NotI and SalI-digested pCMV-SPORT6 vector. The library was constructed at DOE Joint Genome Institute (Walnut Creek, CA).

text: Early embryo

text: Nematostella vectensis

text: SalI

text: NotI

text: CAGF Nematostella vectensis Nemve Early embryo <24hrs


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 at DOE Joint Genome Institute (Walnut Creek, CA).

text: Early embryo

text: Nematostella vectensis

text: SalI

text: NotI

text: CAGG Nematostella vectensis Nemve Early embryo <24hrs


text: DOE Joint Genome Institute

text: live

text: Generic

text: Late embryo

text: Nematostella vectensis

text: SalI

text: NotI

text: CAGH Nematostella vectensis Nemve Late embryo <24hrs


text: DOE Joint Genome Institute

text: live

text: Generic

text: The library was made from oligo(dT)-primed cDNA and cloned into the pCMV-SPORT6.1 vector (Invitrogen Corp., Carlsbad, CA). Library constructed at the Sars International Centre for Marine Molecular Biology and sequencing was done at DOE Joint Genome Institute (Walnut Creek, CA).

text: unfertilized eggs to primary polyps

text: Nematostella vectensis

text: CAGN Nematostella vectensis Nemve mixed stages unfert eggs to primary polyps


text: DOE Joint Genome Institute

text: live

text: Generic

text: The library was made from oligo(dT)-primed cDNA and cloned into the pBluescript SK+ vector (Stratagene, La Jolla, CA). Library is normalized. Library constructed at the Sars International Centre for Marine Molecular Biology and sequencing was done at DOE Joint Genome Institute (Walnut Creek, CA).

text: Nematostella vectensis

text: CAIC Nematostella vectensis Nemve whole embryos normalized


text: University of St Andrews, Bute Medical Building

text: live

text: Generic

text: Anguilla anguilla

text: Anguilla anguilla library (Kalujnaia S)


text: Austrian Research Centers GmbH - ARC

text: live

text: Generic

text: Yungay

text: country: Peru

text: Solanum tuberosum subsp. andigenum

text: leaf

characteristic: tissue-type

text: Solanum tuberosum subsp. andigena leaf


text: Austrian Research Centers GmbH - ARC

text: live

characteristic: INSDC status

text: Generic

text: Yungay

text: country: Peru

text: Solanum caripense

text: leaf

text: Solanum caripense cv. Yungay leaf


text: Austrian Research Centers GmbH - ARC

text: live

text: Generic

text: country: Peru

text: Solanum caripense

text: leaf

text: Solanum caripense leaf


text: DOE Joint Genome Institute

text: live

text: Generic

text: In Vivo Conditions: Solid culture media, BCDA media. The library was made from oligo(dT)-primed cDNA and cloned into the pMCL200_cdna vector. PolyA RNA was primed with 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 pMCL200_cdna vector. Library was created and sequencing done by the DOE Joint Genome Institute (Walnut Creek, CA).

text: Asexual stage, monoic organism

characteristic: lab-host

text: Physcomitrium patens

text: SalI

text: NotI

text: Villersexel 2003

text: Protonema

text: CAYY Physcomitrella patens subsp. patens Villersexel - regular (H)

characteristic: vector


text: DOE Joint Genome Institute

text: live

text: Generic

characteristic: NCBI submission model

text: In Vivo Conditions: Solid culture media, BCDA media. The library was made from oligo(dT)-primed cDNA and cloned into the pMCL200_cdna vector. PolyA RNA was primed with 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 1-2.5kb using 1.1% agarose gel electrophoresis then ligated into NotI and SalI-digested pMCL200_cdna vector. Library was created and sequencing done by the DOE Joint Genome Institute (Walnut Creek, CA).

text: Asexual stage, monoic organism

text: Physcomitrium patens

text: SalI

text: NotI

text: Villersexel 2003

text: Protonema

text: CAYZ Physcomitrella patens subsp. patens Villersexel - regular (L)


text: DOE Joint Genome Institute

text: live

text: Generic

characteristic: NCBI submission package

text: In Vivo Conditions: Solid culture media, BCDA media. 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 at DOE Joint Genome Institute (Walnut Creek, CA).

text: haploid stage, 7 days old

text: DH10B T1R

text: Physcomitrium patens

text: SalI

text: NotI

text: Gransden 2004

text: Protonema

text: CBAW Physcomitrella patens subsp. patens Gransden - dessicated (H)


text: DOE Joint Genome Institute

text: live

text: Generic

text: In Vivo Conditions: Solid culture media, BCDA media. 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 at DOE Joint Genome Institute (Walnut Creek, CA).

text: haploid stage, 7 days old

text: DH10B T1R

text: Physcomitrium patens

text: SalI

text: NotI

text: Gransden 2004

text: Protonema

text: CBAX Physcomitrella patens subsp. patens Gransden - dessicated (L)


text: Veterinary College, Anand Agricultural University

text: live

text: Generic

text: Epithelial Cell

text: Total RNA was isolated from mammary gland of four different physiolgical stages of buffalo (heifer, pregnant, lactating and dry). Differential ESTs were identified from mRNA.

text: Heifer

text: E. coli DH-5alpha

text: Mammary gland

characteristic: organ

text: Bubalus bubalis

text: dT

text: dT

text: Female

characteristic: sex

text: Surti breed

text: Mammary tissue

text: Mammary tissue of buffalo heifer

text: Plasmid


text: live

text: Generic

text: Epithelial Cell

text: dry buffalo

text: E. coli DH-5alpha

text: Mammary gland

text: Bubalus bubalis

characteristic: organism

text: dT

text: dT

text: Female

text: Surti breed

characteristic: strain

text: Mammary tissue

text: Mammary tissue of dry buffalo

text: Plasmid


text: live

text: Generic

text: Epithelial Cell

characteristic: description

text: lactating buffalo

text: E. coli DH-5alpha

text: Mammary gland

text: Bubalus bubalis

text: dT

text: dT

text: Female

text: Surti breed

text: Mammary tissue

text: Mammary tissue of lactating buffalo

characteristic: title

text: Plasmid


characteristic: External Id

text: live

text: Generic

text: Epithelial Cell

characteristic: cell type

text: pregnant

text: E. coli DH-5alpha

text: Mammary gland

text: Bubalus bubalis

text: dT

text: dT

text: Female

text: Surti breed

text: Mammary tissue

text: Mammary tissue of pregnant buffalo

text: Plasmid


characteristic: INSDC center name

text: live

text: Generic

text: Epithelial cells

text: Total RNA was isolated from mammary gland of three different stages of buffalo (zero hours, 48 hours and 96 hours of rbST treatment). Differential ESTs were identified by DDRT-PCR. Differentially Displayed band was cloned and sequenced.

text: lactating buffalo

characteristic: dev-stage

text: E. coli DH-5alpha

text: Bubalus bubalis

text: dT

text: dT

text: female

text: Surti Breed

text: mammary tissue

text: Mammary tissue of lactating female buffalo at 0 hours of rbST treatment

text: Plasmid


characteristic: INSDC first public

characteristic: INSDC last update

text: live

text: Generic

text: Epithelial cells

text: lactating buffalo

text: E. coli DH-5alpha

text: Bubalus bubalis

text: dT

text: dT

text: female

text: Surti Breed

text: mammary tissue

text: Mammary tissue of lactating female buffalo at 48 hours of rbST treatment

text: Plasmid


text: live

characteristic: INSDC status

text: Generic

text: Epithelial cells

text: lactating buffalo

text: E. coli DH-5alpha

text: Bubalus bubalis

text: dT

text: dT

text: female

text: Surti Breed

text: mammary tissue

characteristic: tissue-type

text: Mammary tissue of lactating female buffalo at 96 hours of rbST treatment

text: Plasmid


text: INRA (1064) CNRS (6192) Universite de Nice Sophia Antipolis

text: live

text: Generic

text: Labrador

characteristic: cultivar

text: SSH method was used to isolate gene specifically up-regulated in response to drought in soybean nodule. mRNA were extracted from control plant and from drought stressed (5 days without watering) plant. cDNA obtained after subtraction were cloned into pGEMT vector and sequenced with the T7 primer.

text: mature soybean, six weeks old

text: nodule

text: Glycine max

text: Soybean nodule 5 days after drought library


text: University of California

text: live

text: Generic

characteristic: NCBI submission model

text: Seeds of 524B were from a single plant with a long history of single seed descent (Ehlers)from a California breeding program. The types of tissue were: 1) germinated seed hypocotyl and primary root, 2) germinated seed epicotyl and foliage leaves, 3 & 4) young trifoliate leaves and apical meristem (one well watered, one drought stressed). For germinated seed samples (1 & 2), seeds were surface sterilized 5 minutes in 1% sodium hypochlorite with 0.01% Tween 20, rinsed with water, germinated on sterile filter paper with water containing 100 units/ml nystatin and 50 microgram/ml cefotaxime, incubated 4-5 days in the dark at room temperature (Fenton). For samples 3 & 4, seeds were planted in 3 gallon pots and kept watered in a glasshouse until plants were ~25 cm in height (~3 weeks)(Fenton). For drought stress (sample 4), water was withheld until visible signs of drought stress occurred (reduced growth relative to watered plants, wilted leaves, less green color), generally ~3 weeks. Total RNA was purified using a TRIzol method, except Qiagen RNeasy for hypocotyls & root (Fenton). RNA was pooled to establish one complex RNA sample with a total of 400 microgram, as follows: 133 microgram of hypocotyl & primary root (sample 1), 133 microgram of epicotyl and foliage leaves (sample 2) and 67 microgram each of stressed and unstressed young trifoliate leaves and apical meristem (samples 3 & 4)(Fenton). PolyA RNA was purified using Absolutely mRNA Purification Kit (Stratagene)(Fenton). 1.7 microgram of poly(A) mRNA was used for cDNA synthesis. A primary cDNA library with a titer of 10.9 million cfu (Close) was produced using a SuperScript Plasmid System with Gateway Technology for cDNA Synthesis and Cloning (Invitrogen)(Fenton). Plating and sequencing was performed at the US Department of Energy Joint Genome Institute Sequence Production Facility, Walnut Creek California (Richardson, Lucas, Rokhsar, Wang, Lindquist). The library was plated onto agarose plates containing ampicillin (254 mm plates from Teknova, Hollister, CA) at a density of approximately 1000 colonies per plate. Plates were grown at 37 degrees Celsius for 18 hours, then individual colonies were picked and each used to inoculate a well containing LB media with ampicillin in a 384 well plate (Nunc, Rochester, NY). 384 well plates were grown at 37 degrees Celsius for 18 hours. Plasmid DNA for sequencing was produced by rolling circle amplification (Templiphi, GE Healthcare, Piscataway, NJ). Subclone inserts were sequenced from both ends using primers complementary to the flanking pCMV-SPORT6 vector sequence and Big Dye terminator chemistry and then run on ABI 3730 instruments (ABI, Foster City, CA). Sequencing protocols can be found in Detter et al (2002, Genomics 80:691-698) and at http://www.jgi.doe.gov/sequencing/protocols/index.html. Trace files were downloaded (Close) and then processed (Wanamaker) using the HarvEST pipeline (http://harvest.ucr.edu) to remove vector and cloning oligo sequences and various contaminants, and to trim to a high quality region. Sequences that retained a phred 17 region of at least 100 bases were deposited to GenBank with annotations (Close).

text: Seedling

text: E. coli DH10B

characteristic: lab-host

text: Vigna unguiculata

text: SalI

text: NotI

text: mixed

text: Cowpea 524B Mixed Tissue and Conditions cDNA Library UCRVU04-1

text: Plasmid

characteristic: vector


text: University of California

text: live

text: Generic

characteristic: NCBI submission package

text: Seeds of IT84S-2049 were from a single plant with a long history of single seed descent (Ehlers) from a Nigerian breeding program. The types of tissue were: 1) germinated seed hypocotyl and primary root, 2) germinated seed epicotyl and foliage leaves, 3 & 4) young trifoliate leaves and apical meristem (one well watered, one drought stressed). For germinated seed samples (1 & 2), seeds were surface sterilized 5 minutes in 1% sodium hypochlorite with 0.01% Tween 20, rinsed with water, germinated on sterile filter paper with water containing 100 units/ml nystatin and 50 microgram/ml cefotaxime, incubated 4-5 days in the dark at room temperature (Fenton). For samples 3 & 4, seeds were planted in 3 gallon pots and kept watered in a glasshouse until plants were ~25 cm in height (~3 weeks)(Fenton). For drought stress (sample 4), water was withheld until visible signs of drought stress occurred (reduced growth relative to watered plants, wilted leaves, less green color), generally ~3 weeks. Total RNA was purified using a TRIzol method, except Qiagen RNeasy for hypocotyls & root (Fenton). RNA was pooled to establish one complex RNA sample with a total of 400 microgram, as follows: 133 microgram of hypocotyl & primary root (sample 1), 133 microgram of epicotyl and foliage leaves (sample 2) and 67 microgram each of stressed and unstressed young trifoliate leaves and apical meristem (samples 3 & 4)(Fenton). PolyA RNA was purified using Absolutely mRNA Purification Kit (Stratagene)(Fenton). 3.0 microgram of poly(A) mRNA was used for cDNA synthesis. A primary cDNA library with a titer of 17.6 million cfu (Close) was produced using a SuperScript Plasmid System with Gateway Technology for cDNA Synthesis and Cloning (Invitrogen)(Fenton). Plating and sequencing was performed at the US Department of Energy Joint Genome Institute Sequence Production Facility, Walnut Creek California (Richardson, Lucas, Rokhsar, Wang, Lindquist). The library was plated onto agarose plates containing ampicillin (254 mm plates from Teknova, Hollister, CA) at a density of approximately 1000 colonies per plate. Plates were grown at 37 degrees Celsius for 18 hours, then individual colonies were picked and each used to inoculate a well containing LB media with ampicillin in a 384 well plate (Nunc, Rochester, NY). 384 well plates were grown at 37 degrees Celsius for 18 hours. Plasmid DNA for sequencing was produced by rolling circle amplification (Templiphi, GE Healthcare, Piscataway, NJ). Subclone inserts were sequenced from both ends using primers complementary to the flanking pCMV-SPORT6 vector sequence and Big Dye terminator chemistry and then run on ABI 3730 instruments (ABI, Foster City, CA). Sequencing protocols can be found in Detter et al (2002, Genomics 80:691-698) and at http://www.jgi.doe.gov/sequencing/protocols/index.html. Trace files were downloaded (Close) and then processed (Wanamaker) using the HarvEST pipeline (http://harvest.ucr.edu) to remove vector and cloning oligo sequences and various contaminants, and to trim to a high quality region. Sequences that retained a phred 17 region of at least 100 bases were deposited to GenBank with annotations (Close).

text: Seedling

text: E. coli DH10B

text: Vigna unguiculata

text: SalI

text: NotI

text: mixed

text: Cowpea IT84S-2049 Mixed Tissue and Conditions cDNA Library UCRVU05-2

text: Plasmid


text: University of California

text: live

text: Generic

text: UCR 5301

text: Seeds of UCR 5301 were from a single plant with a long history of single seed descent (Ehlers) from a wild cowpea accession. The types of tissue were: 1) germinated seed hypocotyl and primary root, 2) germinated seed epicotyl and foliage leaves, 3 & 4) young trifoliate leaves and apical meristem (one well watered, one drought stressed). For germinated seed samples (1 & 2), seeds were surface sterilized 5 minutes in 1% sodium hypochlorite with 0.01% Tween 20, rinsed with water, germinated on sterile filter paper with water containing 100 units/ml nystatin and 50 microgram/ml cefotaxime, incubated 4-5 days in the dark at room temperature (Fenton). For samples 3 & 4, seeds were planted in 3 gallon pots and kept watered in a glasshouse until plants were ~25 cm in height (~3 weeks)(Fenton). For drought stress (sample 4), water was withheld until visible signs of drought stress occurred (reduced growth relative to watered plants, wilted leaves, less green color), generally ~3 weeks. Total RNA was purified using a TRIzol method, except Qiagen RNeasy for hypocotyls & root (Fenton). RNA was pooled to establish one complex RNA sample with a total of 500 microgram, as follows: 167 microgram of hypocotyl & primary root (sample 1), 167 microgram of epicotyl and foliage leaves (sample 2) and 87 microgram each of stressed and unstressed young trifoliate leaves and apical meristem (samples 3 & 4)(Fenton). PolyA RNA was purified using PolyATrack mRNA Isolation System IV (Promega)(Fenton). 2.0 microgram of poly(A) mRNA was used for cDNA synthesis. A primary cDNA library with a titer of 25.6 million cfu (Close) was produced using a SuperScript Plasmid System with Gateway Technology for cDNA Synthesis and Cloning (Invitrogen)(Fenton). Plating and sequencing was performed at the US Department of Energy Joint Genome Institute Sequence Production Facility, Walnut Creek California (Richardson, Lucas, Rokhsar, Wang, Lindquist). The library was plated onto agarose plates containing ampicillin (254 mm plates from Teknova, Hollister, CA) at a density of approximately 1000 colonies per plate. Plates were grown at 37 degrees Celsius for 18 hours, then individual colonies were picked and each used to inoculate a well containing LB media with ampicillin in a 384 well plate (Nunc, Rochester, NY). 384 well plates were grown at 37 degrees Celsius for 18 hours. Plasmid DNA for sequencing was produced by rolling circle amplification (Templiphi, GE Healthcare, Piscataway, NJ). Subclone inserts were sequenced from both ends using primers complementary to the flanking pCMV-SPORT6 vector sequence and Big Dye terminator chemistry and then run on ABI 3730 instruments (ABI, Foster City, CA). Sequencing protocols can be found in Detter et al (2002, Genomics 80:691-698) and at http://www.jgi.doe.gov/sequencing/protocols/index.html. Trace files were downloaded (Close) and then processed (Wanamaker) using the HarvEST pipeline (http://harvest.ucr.edu) to remove vector and cloning oligo sequences and various contaminants, and to trim to a high quality region. Sequences that retained a phred 17 region of at least 100 bases were deposited to GenBank with annotations (Close).

text: Seedling

text: E. coli DH10B

text: Vigna unguiculata

characteristic: organism

text: SalI

text: NotI

text: mixed

text: Cowpea UCR 5301 Mixed Tissue and Conditions cDNA Library UCRVU06-1

text: Plasmid


text: University of California

text: live

text: Generic

text: UCR 779

text: Seeds of UCR 779 were from a single plant with a long history of single seed descent (Ehlers) from Botswana. The types of tissue were: 1) germinated seed hypocotyl and primary root, 2) germinated seed epicotyl and foliage leaves, 3 & 4) young trifoliate leaves and apical meristem (one well watered, one drought stressed). For germinated seed samples (1 & 2), seeds were surface sterilized 5 minutes in 1% sodium hypochlorite with 0.01% Tween 20, rinsed with water, germinated on sterile filter paper with water containing 100 units/ml nystatin and 50 microgram/ml cefotaxime, incubated 4-5 days in the dark at room temperature (Fenton). For samples 3 & 4, seeds were planted in 3 gallon pots and kept watered in a glasshouse until plants were ~25 cm in height (~3 weeks)(Fenton). For drought stress (sample 4), water was withheld until visible signs of drought stress occurred (reduced growth relative to watered plants, wilted leaves, less green color), generally ~3 weeks. Total RNA was purified using a TRIzol method, except Qiagen RNeasy for hypocotyls & root (Fenton). RNA was pooled to establish one complex RNA sample with a total of 500 microgram, as follows: 167 microgram of hypocotyl & primary root (sample 1), 167 microgram of epicotyl and foliage leaves (sample 2) and 87 microgram each of stressed and unstressed young trifoliate leaves and apical meristem (samples 3 & 4)(Fenton). PolyA RNA was purified using PolyATrack mRNA Isolation System IV (Promega)(Fenton). 2.0 microgram of poly(A) mRNA was used for cDNA synthesis. A primary cDNA library with a titer of 23 million cfu (Close) was produced using a SuperScript Plasmid System with Gateway Technology for cDNA Synthesis and Cloning (Invitrogen)(Fenton). Plating and sequencing was performed at the US Department of Energy Joint Genome Institute Sequence Production Facility, Walnut Creek California (Richardson, Lucas, Rokhsar, Wang, Lindquist). The library was plated onto agarose plates containing ampicillin (254 mm plates from Teknova, Hollister, CA) at a density of approximately 1000 colonies per plate. Plates were grown at 37 degrees Celsius for 18 hours, then individual colonies were picked and each used to inoculate a well containing LB media with ampicillin in a 384 well plate (Nunc, Rochester, NY). 384 well plates were grown at 37 degrees Celsius for 18 hours. Plasmid DNA for sequencing was produced by rolling circle amplification (Templiphi, GE Healthcare, Piscataway, NJ). Subclone inserts were sequenced from both ends using primers complementary to the flanking pCMV-SPORT6 vector sequence and Big Dye terminator chemistry and then run on ABI 3730 instruments (ABI, Foster City, CA). Sequencing protocols can be found in Detter et al (2002, Genomics 80:691-698) and at http://www.jgi.doe.gov/sequencing/protocols/index.html. Trace files were downloaded (Close) and then processed (Wanamaker) using the HarvEST pipeline (http://harvest.ucr.edu) to remove vector and cloning oligo sequences and various contaminants, and to trim to a high quality region. Sequences that retained a phred 17 region of at least 100 bases were deposited to GenBank with annotations (Close).

text: Seedling

text: E. coli DH10B

text: Vigna unguiculata

text: SalI

text: NotI

text: mixed

text: Cowpea UCR 779 Mixed Tissue and Conditions cDNA Library UCRVU07-1

text: Plasmid


text: University of California

text: live

text: Generic

text: Seeds of IT97K-461-4 were from a single plant with a long history of single seed descent (Ehlers) from a Nigerian breeding program. The types of tissue were: 1) germinated seed hypocotyl and primary root, 2) germinated seed epicotyl and foliage leaves, 3 & 4) young trifoliate leaves and apical meristem (one well watered, one drought stressed). For germinated seed samples (1 & 2), seeds were surface sterilized 5 minutes in 1% sodium hypochlorite with 0.01% Tween 20, rinsed with water, germinated on sterile filter paper with water containing 100 units/ml nystatin and 50 microgram/ml cefotaxime, incubated 4-5 days in the dark at room temperature (Fenton). For samples 3 & 4, seeds were planted in 3 gallon pots and kept watered in a glasshouse until plants were ~25 cm in height (~3 weeks)(Fenton). For drought stress (sample 4), water was withheld until visible signs of drought stress occurred (reduced growth relative to watered plants, wilted leaves, less green color), generally ~3 weeks. Total RNA was purified using a TRIzol method, except Qiagen RNeasy for hypocotyls & root (Fenton). RNA was pooled to establish one complex RNA sample with a total of 500 microgram, as follows: 167 microgram of hypocotyl & primary root (sample 1), 167 microgram of epicotyl and foliage leaves (sample 2) and 87 microgram each of stressed and unstressed young trifoliate leaves and apical meristem (samples 3 & 4)(Fenton). PolyA RNA was purified using PolyATrack mRNA Isolation System IV (Promega)(Fenton). 2.0 microgram of poly(A) mRNA was used for cDNA synthesis. A primary cDNA library with a titer of 20 million cfu (Close) was produced using a SuperScript Plasmid System with Gateway Technology for cDNA Synthesis and Cloning (Invitrogen)(Fenton). Plating and sequencing was performed at the US Department of Energy Joint Genome Institute Sequence Production Facility, Walnut Creek California (Richardson, Lucas, Rokhsar, Wang, Lindquist). The library was plated onto agarose plates containing ampicillin (254 mm plates from Teknova, Hollister, CA) at a density of approximately 1000 colonies per plate. Plates were grown at 37 degrees Celsius for 18 hours, then individual colonies were picked and each used to inoculate a well containing LB media with ampicillin in a 384 well plate (Nunc, Rochester, NY). 384 well plates were grown at 37 degrees Celsius for 18 hours. Plasmid DNA for sequencing was produced by rolling circle amplification (Templiphi, GE Healthcare, Piscataway, NJ). Subclone inserts were sequenced from both ends using primers complementary to the flanking pCMV-SPORT6 vector sequence and Big Dye terminator chemistry and then run on ABI 3730 instruments (ABI, Foster City, CA). Sequencing protocols can be found in Detter et al (2002, Genomics 80:691-698) and at http://www.jgi.doe.gov/sequencing/protocols/index.html. Trace files were downloaded (Close) and then processed (Wanamaker) using the HarvEST pipeline (http://harvest.ucr.edu) to remove vector and cloning oligo sequences and various contaminants, and to trim to a high quality region. Sequences that retained a phred 17 region of at least 100 bases were deposited to GenBank with annotations (Close).

text: Seedling

text: E. coli DH10B

text: Vigna unguiculata

text: SalI

text: NotI

text: mixed

text: Cowpea IT97K-461-4 Mixed Tissue and Conditions cDNA Library UCRVU08-1

text: Plasmid


text: University of California

text: live

text: Generic

text: UCR 707

text: Seeds of UCR 707 were from a single plant with a long history of single seed descent (Ehlers) from Kenya. The types of tissue were: 1) germinated seed hypocotyl and primary root, 2) germinated seed epicotyl and foliage leaves, 3 & 4) young trifoliate leaves and apical meristem (one well watered, one drought stressed). For germinated seed samples (1 & 2), seeds were surface sterilized 5 minutes in 1% sodium hypochlorite with 0.01% Tween 20, rinsed with water, germinated on sterile filter paper with water containing 100 units/ml nystatin and 50 microgram/ml cefotaxime, incubated 4-5 days in the dark at room temperature (Fenton). For samples 3 & 4, seeds were planted in 3 gallon pots and kept watered in a glasshouse until plants were ~25 cm in height (~3 weeks)(Fenton). For drought stress (sample 4), water was withheld until visible signs of drought stress occurred (reduced growth relative to watered plants, wilted leaves, less green color), generally ~3 weeks. Total RNA was purified using a TRIzol method, except Qiagen RNeasy for hypocotyls & root (Fenton). RNA was pooled to establish one complex RNA sample with a total of 500 microgram, as follows: 167 microgram of hypocotyl & primary root (sample 1), 167 microgram of epicotyl and foliage leaves (sample 2) and 87 microgram each of stressed and unstressed young trifoliate leaves and apical meristem (samples 3 & 4)(Fenton). PolyA RNA was purified using PolyATrack mRNA Isolation System IV (Promega)(Fenton). 2.0 microgram of poly(A) mRNA was used for cDNA synthesis. A primary cDNA library with a titer of 30 million cfu (Close) was produced using a SuperScript Plasmid System with Gateway Technology for cDNA Synthesis and Cloning (Invitrogen)(Fenton). Plating and sequencing was performed at the US Department of Energy Joint Genome Institute Sequence Production Facility, Walnut Creek California (Richardson, Lucas, Rokhsar, Wang, Lindquist). The library was plated onto agarose plates containing ampicillin (254 mm plates from Teknova, Hollister, CA) at a density of approximately 1000 colonies per plate. Plates were grown at 37 degrees Celsius for 18 hours, then individual colonies were picked and each used to inoculate a well containing LB media with ampicillin in a 384 well plate (Nunc, Rochester, NY). 384 well plates were grown at 37 degrees Celsius for 18 hours. Plasmid DNA for sequencing was produced by rolling circle amplification (Templiphi, GE Healthcare, Piscataway, NJ). Subclone inserts were sequenced from both ends using primers complementary to the flanking pCMV-SPORT6 vector sequence and Big Dye terminator chemistry and then run on ABI 3730 instruments (ABI, Foster City, CA). Sequencing protocols can be found in Detter et al (2002, Genomics 80:691-698) and at http://www.jgi.doe.gov/sequencing/protocols/index.html. Trace files were downloaded (Close) and then processed (Wanamaker) using the HarvEST pipeline (http://harvest.ucr.edu) to remove vector and cloning oligo sequences and various contaminants, and to trim to a high quality region. Sequences that retained a phred 17 region of at least 100 bases were deposited to GenBank with annotations (Close).

text: Seedling

text: E. coli DH10B

text: Vigna unguiculata

text: SalI

text: NotI

text: mixed

text: Cowpea UCR 707 Mixed Tissue and Conditions cDNA Library UCRVU09-1

text: Plasmid


text: University of California

text: live

text: Generic

text: PI 418979

text: Seeds of PI 418979 were from a single plant with a long history of single seed descent (Ehlers) from China. The types of tissue were: 1) germinated seed hypocotyl and primary root, 2) germinated seed epicotyl and foliage leaves, 3 & 4) young trifoliate leaves and apical meristem (one well watered, one drought stressed). For germinated seed samples (1 & 2), seeds were surface sterilized 5 minutes in 1% sodium hypochlorite with 0.01% Tween 20, rinsed with water, germinated on sterile filter paper with water containing 100 units/ml nystatin and 50 microgram/ml cefotaxime, incubated 4-5 days in the dark at room temperature (Fenton). For samples 3 & 4, seeds were planted in 3 gallon pots and kept watered in a glasshouse until plants were ~25 cm in height (~3 weeks)(Fenton). For drought stress (sample 4), water was withheld until visible signs of drought stress occurred (reduced growth relative to watered plants, wilted leaves, less green color), generally ~3 weeks. Total RNA was purified using a TRIzol method, except Qiagen RNeasy for hypocotyls & root (Fenton). RNA was pooled to establish one complex RNA sample with a total of 500 microgram, as follows: 167 microgram of hypocotyl & primary root (sample 1), 167 microgram of epicotyl and foliage leaves (sample 2) and 87 microgram each of stressed and unstressed young trifoliate leaves and apical meristem (samples 3 & 4)(Fenton). PolyA RNA was purified using PolyATrack mRNA Isolation System IV (Promega)(Fenton). 2.0 microgram of poly(A) mRNA was used for cDNA synthesis. A primary cDNA library with a titer of 15 million cfu (Close) was produced using a SuperScript Plasmid System with Gateway Technology for cDNA Synthesis and Cloning (Invitrogen)(Fenton). Plating and sequencing was performed at the US Department of Energy Joint Genome Institute Sequence Production Facility, Walnut Creek California (Richardson, Lucas, Rokhsar, Wang, Lindquist). The library was plated onto agarose plates containing ampicillin (254 mm plates from Teknova, Hollister, CA) at a density of approximately 1000 colonies per plate. Plates were grown at 37 degrees Celsius for 18 hours, then individual colonies were picked and each used to inoculate a well containing LB media with ampicillin in a 384 well plate (Nunc, Rochester, NY). 384 well plates were grown at 37 degrees Celsius for 18 hours. Plasmid DNA for sequencing was produced by rolling circle amplification (Templiphi, GE Healthcare, Piscataway, NJ). Subclone inserts were sequenced from both ends using primers complementary to the flanking pCMV-SPORT6 vector sequence and Big Dye terminator chemistry and then run on ABI 3730 instruments (ABI, Foster City, CA). Sequencing protocols can be found in Detter et al (2002, Genomics 80:691-698) and at http://www.jgi.doe.gov/sequencing/protocols/index.html. Trace files were downloaded (Close) and then processed (Wanamaker) using the HarvEST pipeline (http://harvest.ucr.edu) to remove vector and cloning oligo sequences and various contaminants, and to trim to a high quality region. Sequences that retained a phred 17 region of at least 100 bases were deposited to GenBank with annotations (Close).

text: Seedling

text: E. coli DH10B

text: Vigna unguiculata

text: SalI

text: NotI

text: mixed

text: Cowpea PI 418979 Mixed Tissue and Conditions cDNA Library UCRVU10-1

text: Plasmid


text: University of California

text: live

text: Generic

text: UCR 41

text: Seeds of UCR 41 were from a single plant with a long history of single seed descent (Ehlers) from Nicaragua. The types of tissue were: 1) germinated seed hypocotyl and primary root, 2) germinated seed epicotyl and foliage leaves, 3 & 4) young trifoliate leaves and apical meristem (one well watered, one drought stressed). For germinated seed samples (1 & 2), seeds were surface sterilized 5 minutes in 1% sodium hypochlorite with 0.01% Tween 20, rinsed with water, germinated on sterile filter paper with water containing 100 units/ml nystatin and 50 microgram/ml cefotaxime, incubated 4-5 days in the dark at room temperature (Fenton). For samples 3 & 4, seeds were planted in 3 gallon pots and kept watered in a glasshouse until plants were ~25 cm in height (~3 weeks)(Fenton). For drought stress (sample 4), water was withheld until visible signs of drought stress occurred (reduced growth relative to watered plants, wilted leaves, less green color), generally ~3 weeks. Total RNA was purified using a TRIzol method, except Qiagen RNeasy for hypocotyls & root (Fenton). RNA was pooled to establish one complex RNA sample with a total of 500 microgram, as follows: 167 microgram of hypocotyl & primary root (sample 1), 167 microgram of epicotyl and foliage leaves (sample 2) and 87 microgram each of stressed and unstressed young trifoliate leaves and apical meristem (samples 3 & 4)(Fenton). PolyA RNA was purified using PolyATrack mRNA Isolation System IV (Promega)(Fenton). 2.0 microgram of poly(A) mRNA was used for cDNA synthesis. A primary cDNA library with a titer of 18 million cfu (Close) was produced using a SuperScript Plasmid System with Gateway Technology for cDNA Synthesis and Cloning (Invitrogen)(Fenton). Plating and sequencing was performed at the US Department of Energy Joint Genome Institute Sequence Production Facility, Walnut Creek California (Richardson, Lucas, Rokhsar, Wang, Lindquist). The library was plated onto agarose plates containing ampicillin (254 mm plates from Teknova, Hollister, CA) at a density of approximately 1000 colonies per plate. Plates were grown at 37 degrees Celsius for 18 hours, then individual colonies were picked and each used to inoculate a well containing LB media with ampicillin in a 384 well plate (Nunc, Rochester, NY). 384 well plates were grown at 37 degrees Celsius for 18 hours. Plasmid DNA for sequencing was produced by rolling circle amplification (Templiphi, GE Healthcare, Piscataway, NJ). Subclone inserts were sequenced from both ends using primers complementary to the flanking pCMV-SPORT6 vector sequence and Big Dye terminator chemistry and then run on ABI 3730 instruments (ABI, Foster City, CA). Sequencing protocols can be found in Detter et al (2002, Genomics 80:691-698) and at http://www.jgi.doe.gov/sequencing/protocols/index.html. Trace files were downloaded (Close) and then processed (Wanamaker) using the HarvEST pipeline (http://harvest.ucr.edu) to remove vector and cloning oligo sequences and various contaminants, and to trim to a high quality region. Sequences that retained a phred 17 region of at least 100 bases were deposited to GenBank with annotations (Close).

text: Seedling

text: E. coli DH10B

text: Vigna unguiculata

text: SalI

text: NotI

text: mixed

text: Cowpea UCR 41 Mixed Tissue and Conditions cDNA Library UCRVU11-1

text: Plasmid


text: University of California

text: live

text: Generic

text: UCR 2563

text: Seeds of UCR 2563 were from a single plant with a long history of single seed descent (Ehlers)from Brazil. The types of tissue were: 1) germinated seed hypocotyl and primary root, 2) germinated seed epicotyl and foliage leaves, 3 & 4) young trifoliate leaves and apical meristem (one well watered, one drought stressed). For germinated seed samples (1 & 2), seeds were surface sterilized 5 minutes in 1% sodium hypochlorite with 0.01% Tween 20, rinsed with water, germinated on sterile filter paper with water containing 100 units/ml nystatin and 50 microgram/ml cefotaxime, incubated 4-5 days in the dark at room temperature (Fenton). For samples 3 & 4, seeds were planted in 3 gallon pots and kept watered in a glasshouse until plants were ~25 cm in height (~3 weeks)(Fenton). For drought stress (sample 4), water was withheld until visible signs of drought stress occurred (reduced growth relative to watered plants, wilted leaves, less green color), generally ~3 weeks. Total RNA was purified using a TRIzol method, except Qiagen RNeasy for hypocotyls & root (Fenton). RNA was pooled to establish one complex RNA sample with a total of 500 microgram, as follows: 167 microgram of hypocotyl & primary root (sample 1), 167 microgram of epicotyl and foliage leaves (sample 2) and 87 microgram each of stressed and unstressed young trifoliate leaves and apical meristem (samples 3 & 4)(Fenton). PolyA RNA was purified using PolyATrack mRNA Isolation System IV (Promega)(Fenton). 2.0 microgram of poly(A) mRNA was used for cDNA synthesis. A primary cDNA library with a titer of 15.7 million cfu (Close) was produced using a SuperScript Plasmid System with Gateway Technology for cDNA Synthesis and Cloning (Invitrogen)(Fenton). Plating and sequencing was performed at the US Department of Energy Joint Genome Institute Sequence Production Facility, Walnut Creek California (Richardson, Lucas, Rokhsar, Wang, Lindquist). The library was plated onto agarose plates containing ampicillin (254 mm plates from Teknova, Hollister, CA) at a density of approximately 1000 colonies per plate. Plates were grown at 37 degrees Celsius for 18 hours, then individual colonies were picked and each used to inoculate a well containing LB media with ampicillin in a 384 well plate (Nunc, Rochester, NY). 384 well plates were grown at 37 degrees Celsius for 18 hours. Plasmid DNA for sequencing was produced by rolling circle amplification (Templiphi, GE Healthcare, Piscataway, NJ). Subclone inserts were sequenced from both ends using primers complementary to the flanking pCMV-SPORT6 vector sequence and Big Dye terminator chemistry and then run on ABI 3730 instruments (ABI, Foster City, CA). Sequencing protocols can be found in Detter et al (2002, Genomics 80:691-698) and at http://www.jgi.doe.gov/sequencing/protocols/index.html. Trace files were downloaded (Close) and then processed (Wanamaker) using the HarvEST pipeline (http://harvest.ucr.edu) to remove vector and cloning oligo sequences and various contaminants, and to trim to a high quality region. Sequences that retained a phred 17 region of at least 100 bases were deposited to GenBank with annotations (Close).

text: Seedling

text: E. coli DH10B

text: Vigna unguiculata

text: SalI

text: NotI

text: mixed

text: Cowpea UCR 2563 Mixed Tissue and Conditions cDNA Library UCRVU12-2

text: Plasmid


text: Massachusetts Institute Of Technology

text: live

text: Generic

text: Whole animal

text: 3' RACE product for ceh-30 (C33D12.7) from total RNA of a mixed-stage population of wild-type (N2) C. elegans. Reverse transcription performed using the Gibco 'AP' primer. Primary PCR done using a gene-specific primer whose sequence is given in the comments of the EST sequence file and the Gibco 'AUAP' primer. Product was reamplified using a second gene-specific primer whose sequence is given in the comments of the EST sequence file and the Gibco AUAP primer. Products were ligated into pGEM T-Easy. Individual transformants were picked, miniprepped, and sequenced using SP6 and T7 primers. This sequence file has been trimmed to remove primer and vector sequences.

text: Mixed stage population

text: Whole animal

characteristic: organ

text: Caenorhabditis elegans

text: Hermaphrodite

characteristic: sex

characteristic: strain

text: Whole animal

text: N2 total RNA 3' RACE

text: Plasmid

text: pGEM T-Easy


characteristic: External Id

text: Massachusetts Institute Of Technology

text: live

text: Generic

text: Whole animal

text: 5' RACE product for ceh-30 (C33D12.7) from total RNA of a mixed stage population of wild-type (N2) C. elegans. Reverse transcription performed using a gene-specific primer whose sequence is given in the comments of the EST sequence file, tailed using terminal transferase with dCTP, and amplified by PCR using a second gene-specific primer whose sequence is given in the comments of the EST sequence file and the Gibco Abridged Anchor primer. Products were ligated into pGEM T-Easy. Individual transformants were picked, miniprepped, and sequenced using SP6 and T7 primers. This sequence file has been trimmed to remove nucleotides added by terminal transferase and to remove primer and vector sequences.

characteristic: description

text: Mixed stage population

text: Whole animal

text: Caenorhabditis elegans

text: Hermaphrodite

text: Whole animal

text: N2 Total RNA 5' RACE

characteristic: title

text: Plasmid

text: pGEM T-Easy


text: Massachusetts Institute Of Technology

characteristic: INSDC center name

text: live

text: Generic

text: Whole animal

characteristic: cell type

text: RNA Ligase-mediated 5' RACE (RLM-RACE) product for ceh-30 (C33D12.7) from total RNA of a mixed stage population of wild-type (N2) C. elegans. Total RNA was dephosphorylated, the phosphorylase was inactivated, the mRNA was decapped, and a DNA-RNA hybrid oligonucleotide linker was ligated to the phosphorylated 5' ends of processed mRNAs that had been rendered available by decapping. Reverse transcription performed using a gene-specific primer whose sequence is given in the comments of the EST sequence file and amplified by PCR using this same gene-specific primer and a primer specific to the oligonucleotide linker. Products were reamplified using a second gene-specific primer whose sequence is given in the comments of the EST sequence file and a second, nested primer specific to the oligonucleotide linker. Products were ligated into pGEM T-Easy. Individual transformants were picked, miniprepped, and sequenced using SP6 and T7 primers. This sequence file has been trimmed to remove primer and vector sequences.

text: Mixed stage population

characteristic: dev-stage

text: Whole animal

text: Caenorhabditis elegans

text: Hermaphrodite

text: Whole animal

text: N2 Total RNA RLM-RACE

text: Plasmid

text: pGEM T-Easy


text: Massachusetts Institute Of Technology

characteristic: INSDC first public

characteristic: INSDC last update

text: live

characteristic: INSDC status

text: Generic

text: Whole animal

text: RNA Ligase-mediated 5' RACE (RLM-RACE) product for ceh-30 (C33D12.7) from total RNA of a mixed stage population of him-5(e1467) (CB1467) C. elegans. Total RNA was dephosphorylated, the phosphorylase was inactivated, the mRNA was decapped, and a DNA-RNA hybrid oligonucleotide linker was ligated to the phosphorylated 5' ends of processed mRNAs that had been rendered available by decapping. Reverse transcription performed using a gene-specific primer whose sequence is given in the comments of the EST sequence file and amplified by PCR using this same gene-specific primer and a primer specific to the oligonucleotide linker. Products were reamplified using a second gene-specific primer whose sequence is given in the comments of the EST sequence file and a second, nested primer specific to the oligonucleotide linker. Products were ligated into pGEM T-Easy. Individual transformants were picked, miniprepped, and sequenced using SP6 and T7 primers. This sequence file has been trimmed to remove primer and vector sequences.

text: Mixed stage population

text: Whole animal

text: Caenorhabditis elegans

text: Mixed males and hermaphrodites

text: Whole animal

text: CB1467 Total RNA RLM-RACE

text: Plasmid

text: pGEM T-Easy


text: Massachusetts Institute Of Technology

text: live

text: Generic

characteristic: NCBI submission model

text: Whole animal

text: ceh-30 (C33D12.7) PCR product from Zheng Zhou's C. elegans embryonic Two-Hybrid library. PCR was performed using a gene-specific primer whose sequence is given in the comments of the EST sequence file and a primer specific to the library vector. Products were reamplified using a second gene-specific primer whose sequence is given in the comments of the EST sequence file and a second, nested primer specific to the library vector. Products were ligated into pGEM T-Easy. Individual transformants were picked, miniprepped, and sequenced using SP6 and T7 primers. This sequence file has been trimmed to remove primer and vector sequences.

text: Mixed embryos

text: Whole animal

text: Caenorhabditis elegans

text: Hermaphrodite

text: Whole animal

characteristic: tissue-type

text: WE 2-hyb library

text: Plasmid