characteristic: INSDC center name

text: live

text: Generic

text: XL1-Blue MRF'

text: Sus scrofa

text: Xho I

text: EcoR I

text: illeum

text: rill

text: plasmid

text: pBluescript II SK (+)


status: PUBLIC

characteristic: External Id

text: Institute of Hydrobiology, Chinese Academy of Sciences

characteristic: INSDC first public

characteristic: INSDC last update

text: live

characteristic: INSDC status

text: Generic

characteristic: NCBI submission model

characteristic: NCBI submission package

text: Zebrafish (Danio rerio) as nuclear donor and the enucleated Gobiocypris rarus eggs as recipient. SSH cDNA plasmid library was carried out using a PCR-SelectTM Cdna subtraction kit (Clontech, CA, USA). After the secondary PCR, the products were inserted into pGEMT Vector (Promega, USA).

characteristic: description

text: 50% -epiboly stage

tag: attribute

characteristic: dev-stage

text: DH 5A E.coli

characteristic: lab-host

text: Danio rerio

characteristic: organism

text: AB

characteristic: strain

text: early gastrula embryos

characteristic: tissue-type

text: Gobiocypris rarus and zebrafish (Danio rerio) suppression subtractive hybridization (SSH) cDNA library

characteristic: title

text: Plasmid

text: pGEMT

characteristic: vector


text: Washington University School of Medicine

text: live

text: Generic

text: mRNA extracted with QuickPrep kit (Pharmacia). 5 ug mRNA used to prepare cDNA. Fractionated cDNA cloned into Uni-ZAP XR vector with Eco RI at 5' end and Xho I at 3' end. Library excised and now in pBluescript SK. The library was provided by Catherine Lilley (C.J.Lilley@leeds.ac.uk) and Peter Urwin (P.E.Urwin@leeds.ac.uk), Plant Nematology Group, University of Leeds, UK. Sequencing by: Washington University Genome Sequencing Center, St. Louis, MO.

text: young adult

text: Globodera pallida

text: 5': EcoRI

text: 3': Xho I

text: female

characteristic: sex

text: Globodera pallida Pa2 female

text: phagemid Amp resistant

text: Lambda uni-ZAP XR


text: Chinese National Human Genome Center at Shanghai

text: live

text: Generic

text: miracidium

text: Schistosoma japonicum

text: Schistosoma japonicum miracidium cDNA


text: INRA

text: live

text: Generic

text: Funding for construction and subtraction of the cDNA library was provided to J. P. Renard (BDR, INRA). Funding for EST sequencing was provided by a Genanimal contract from FNS-Ministere de la Recherche (INRA, Num: A02085). The cDNA library was constructed, normalized and subtracted by Dr. A. Hernandez (W.M. Keck Center, University of Illinois Urbana-Champaign) and Dr O. Sandra (BDR, INRA) as described in Bonaldo et al.(1996), Genome Research 6, 791-806. The BCBP library is a normalized library of endometrium (caruncular and inter-caruncular regions) subtracted with previously sequenced clones from bovine placenta (BP), bovine embryo (BE) and bovine spleen (BS) libraries produced in the laboratory of Dr. H.A. Lewin (University of Illinois Urbana-Champaign). The double stranded cDNA was size selected (more than 450 bp). Size selected cDNA was ligated to EcoRI adaptors, digested with NotI then directionally cloned into EcoRI-NotI digested pBluescript II SK(+) phagemid vector. Insert size of the bcbp was between 600-3500 bp.

text: Bos taurus

text: caruncular and inter-caruncular regions from day 20, day 32 and day 64 pregnant cows, cows displaying embryo mortality, day 20 cyclic cows

text: Normalized and Subtracted bovine endometrium tissues (bcbp)

text: pBluescript II SK(+)


text: live

text: Generic

text: Total RNA was prepped from G217B yeast (3-day culture grown in Bill Goldmans's lab 10/1998). The RNA was prepped using a Dynal bead protocol. The cDNA synthesis was done at UCSF (Gibco kit) that allowed directional cloning--the 3' end of the cDNAs have a NotI site (site was added in as part of the the oligodT primer) and the 5' end of the cDNAs have a SalI site (synthesized cDNAs were ligated to Sal overhang primers (so no Sal digestion of the cDNAs was required) and then digested with NotI (an infrequent cutter, so chances of cutting within the cDNA sequence itself are low). cDNAs were cloned into the SalI and NotI sites of the shuttle vector pAS209 (this is a S. cerevisiae vector that ends up expressing cDNAs downstream of the inducible GAL1-10 promoter in Saccharomyces).

text: Yeast

text: Histoplasma capsulatum

text: NotI

text: SalI

text: plasmid (ampicillin resistant)


text: live

text: Generic

text: Histoplasma G217B cDNA library.

text: Histoplasma capsulatum

text: NotI

text: SalI

text: plasmid (ampicillin resistant)


text: live

text: Generic

text: polyA mRNA was captured with oligo d(T) magnetic beads and eluted. AAGCAGTGGTAACAACGCAGAGTACTTTTTTTTTTTTTTTVN was used for priming reverse transcription in the presence of SuperScript II RTase. The 5' primer (5' GCGGCCGCCTAATACGACTCACTATAGGG) annealed to cytosine nucleosides added by terminal transferase to the 5' end of the transcript. Amplification during the PCR used primers 5' AAGCAGTGGTAACAACGCAGA and 5' GGCCGCTAATACGACTCACTATAGGG and Clone-TECH Taq DNA polymerase. PCR products between 500 and 2,000 bp were size-fractioned from 0.7% LE/TAE agarose and cloned into the Topo TA site of Invitrogen plasmid vector pCR4-TOPO.

text: Mycelia

text: Histoplasma capsulatum

text: EcoRI flanks TA cloning site

text: EcoRI flanks TA cloning site

text: plasmid (ampicillin and kanamycin resistant)


text: live

text: Generic

text: Yeast

text: Histoplasma capsulatum

text: EcoRI flanks TA cloning site

text: EcoRI flanks TA cloning site


text: live

text: Generic

text: GoldRush

characteristic: cultivar

text: Total RNA was extracted from freeze dried leaf tissue Stage III, using the method described by Wang and Vodkin (Plant Molecular Biology Reporter 12:132-145, 1994). Poly(A)+ mRNA was isolated from total RNA using the PolyATtract mRNA Isolation system III (Promega). The library was prepared using the Stratagene pBluescript II XR cDNA library construction kit. Complementary DNA was synthesized from mRNA using a Poly (dT) sequence with a XhoI restriction site. EcoRI adapters were ligated to the blunt ended cDNA fragments followed by restriction with XhoI. The cDNA insert is protected from XhoI digestion via methylation during first strand cDNA synthesis. The cDNA fragments were directionally cloned into the EcoRI-XhoI restriction site of the pBluescript vector. The ligated cDNA fragments were transformed into E. coli ElectroMax DH10B host cells. Transformation efficiency: 1.0E+07 Average insert Size by PCR: 1283 bp

text: DH10B ampicillin resistant

text: Malus domestica

text: Xho I

text: Eco RI

text: Leaf

text: Phagemid (ampicillin resistant)

text: pBluescript II SK (+)


text: Dept. of Forest Ecology, University of Helsinki

characteristic: INSDC center name

text: live

text: Generic

text: liquid culture

text: Phlebiopsis gigantea

text: Phlebiopsis gigantea (GIGA)

text: pDNR-LIB


characteristic: External Id

text: UMR INRA 1088/CNRS 5184/U. Bourgogne PME, Inra

text: live

text: Generic

text: Medicago truncatula

text: roots

text: Medicago truncatula J5 roots


text: Institute of Botany

characteristic: INSDC first public

text: live

text: Generic

text: 9-week old mycelium grown on 1/2 MMN agar

text: Amanita muscaria

text: Strain 83

text: SSH lib of A. muscaria with Streptomyces 505

text: Plasmid

text: PCR 2.1


text: The Institute for Genomic Research

characteristic: INSDC last update

text: live

text: Generic

text: Plasmodium vivax field isolate cDNA library made in lambda TriplEx2. Inserts cloned unidirectionally in the Sfi 1A and Sfi 1B sites. Mass excision of library produced inserts in pTriplEx2 plasmid. Inserts sequenced from either 5' or 3' end using TriplEx2 sequencing primer or polydT 24 bp primer respectively.

text: Plasmodium vivax

text: Sfi 1A

text: Sfi 1B

text: Field isolate

text: Field isolate cDNA library

text: Phagemid

text: Lambda TriplEx2


text: Department of Primary Industries

text: live

characteristic: INSDC status

text: Generic

text: SMART PCR cDNA synthesis kit (Clontech, USA). pGEM-T Easy vector system I (Promega, USA). E.coli XL10- Gold ultracompetent cells (Stratagene, USA).

characteristic: isolate

text: Epichloe festucae

text: hyphae (liquid culture)


text: Department of Primary Industries

text: live

text: Generic

characteristic: NCBI submission model

characteristic: NCBI submission package

text: Epichloe coenophiala

text: hyphae (liquid culture)


text: Department of Primary Industries

text: live

text: Generic

characteristic: description

text: Epichloe coenophiala

text: hyphae (solid culture)


text: Department of Primary Industries

text: live

text: Generic

text: Epichloe festucae var. lolii

text: hyphae (solid culture)


text: Department of Primary Industries

text: live

text: Generic

text: Epichloe festucae var. lolii

text: hyphae (liquid culture)


text: Department of Primary Industries

text: live

text: Generic

text: Lolium perenne

text: seed germination library


text: Department of Primary Industries

text: live

text: Generic

text: Lolium perenne

text: subtractive library-12 weeks infected/uninfected ryegrass leaf sheaths


text: Gyeongsang National University

text: live

text: Generic

text: Jangwon

text: XL-1 Blue

characteristic: lab-host

text: Brassica rapa subsp. pekinensis

text: Sal I

text: Not I

text: Chinese cabbage seedling library

text: Plasmid


text: The Canadian Potato Genome Project - BioAtlantech

text: live

text: Generic

text: Shepody

text: supplier: Developmental series. Plants from pathogen-free Solanum tuberosum var. Shepody, clone 1756, nuclear stock were grown in a screenhouse under natural conditions. Individual stolon, swollen stolon, tuber skin and mature tubers (stored for one month at 9oC, 95% relative humidity), were collected and used for individual cDNA library syntheses. The cDNA purification column end fractions for each of these individual libraries were pooled and size-fractionated on an agarose gel. cDNAs between 100 bp and just under 500 bp were excised, purified and ligated into the cloning vector.

text: Solanum tuberosum

characteristic: organism

text: EcoRI

text: XhoI

text: Assorted Stolon/Tuber Developmental Series

text: Low Molecular Weight

text: pBluescript II SK(+) XR


text: Marine Biological Laboratory

text: live

text: Generic

text: adult

characteristic: dev-stage

text: Argopecten irradians

text: gonad

text: Bay Scallop gonad ZAP Express Library

text: pBK-CMV


text: University of Guelph

text: live

text: Generic

text: OAC Rex

text: Based on homology searches, pairs of gene-specific primers were designed to clone phenylpropanoid pathway genes in bean (Phaseolus vulgaris L.). PCR was performed with cDNA from a white bean variety OAC Rex. The resultant fragments were cloned (TOPO TA, Invitrogen) and plasmid DNA containing insert sequenced in both directions.

text: six day old seedling

text: Phaseolus vulgaris

text: whole seedling

characteristic: tissue-type

text: Phaseolus vulgaris (variety OAC Rex) / cDNA

text: pCR 2.1-TOPO


text: Chinese National Human Genome Center at Shanghai

text: live

text: Generic

text: Schistosoma japonicum

text: Schistosoma japonicum reverse cDNA

characteristic: title


text: National Institute of Genetics

text: live

text: Generic

text: country: Japan

text: 11.5 dpc

text: Mus musculus

text: presomitic mesoderm

text: Mus musculus presomitic mesoderm 11.5 dpc


text: Industrial Technology Center of Okayama Prefecture

text: live

text: Generic

text: Monascus pilosus

characteristic: strain

text: Monascus pilosus IFO4520


text: live

text: Generic

text: The library was constructed and normalized according Bonaldo and Soares, Genome Analysis - A Laboratory Manual, Vol. 2: 49 - 157. Inserts are directionally cloned into the EcoRI and NotI sites. The library tag located between the NotI site and the polyA tail is TAGGTTACAC.

text: Rattus norvegicus

text: Vestibular end organs, primary afferent neurons

text: Wackym-Soares normalized rat vestibular cDNA library

text: Plasmid

characteristic: vector


text: live

text: Generic

text: schistosomula

text: Schistosoma japonicum

text: SJS


text: IFREMER-CNRS-Universite de Montpellier 2

text: live

text: Generic

text: hemocytes

characteristic: cell type

text: Subtracted library was constructed with the PCR-select cDNA subtraction kit (Clontech). The subtraction was realised between mRNAs of hemocytes from uninfected shrimps (Driver) and from shrimps surviving to a Vibrio penaeicida infection (Tester: mix of samples collected at 12, 24 and 48 hours post-infection). The cDNA fragments obtained were cloned into pCR(r)2.1-TOPO cloning vector using TOPO TA cloning kit (Invitrogen) and transformed into Escherichia coli TOP10 One Shot Chemically Competent cells (Invitrogen).

text: Adult

text: Penaeus stylirostris

text: L. stylirostris hemocytes subtracted Library

text: pCR(r)2.1-TOPO cloning vector from TOPO TA cloning kit (Invitrogen)


text: National Institute of Agricultural Biotechnology

characteristic: INSDC center name

text: live

text: Generic

text: Tea

characteristic: cultivar

text: PCR-select cDNA subtractive library

text: E.coli DH5-Alpha

text: Camellia sinensis

text: EcoRI

text: leaf

text: Subtractive cDNA library specific for young leaves of tea

text: Plasmid

text: pGEM-T Easy


characteristic: External Id

characteristic: INSDC first public

text: live

text: Generic

text: Jinjachukmyun

text: PCR-select cDNA subtractive library

text: 30days after germination

text: E.coli DH5-Alpha

text: Lactuca sativa

text: EcoRI

text: leaf

text: Subtractive cDNA library specific for UV-B irradiated lettuce leaves

text: Plasmid

text: pGEM-T Easy


text: Department of Botany & Plant Sciences, University of California

characteristic: INSDC last update

text: live

characteristic: INSDC status

text: Generic

text: Pomeroy OP

text: Plants were grown in the greenhouse at University of California, Riverside. The scion was a open-pollinated (very probably selfed) seedling of Poncirus trifoliata cv Pomeroy that was selected as homozygous for the Ctv resistance gene. The rootstock was sweet orange infected with citrus tristeza virus (CTV) isolate T514 over 1 year before sampling (CTV infects sweet orange, but not genotypes carrying the Ctv resistance gene). Shoots 10-30 cm long were harvested in October 2000, and the green phloem (bark) was removed and frozen quickly in dry ice. Total RNA was extracted using TRIZOL reagent (Gibco). Poly(A) RNA was purified, a cDNA library was made, and 0.5 million primary lambda cDNA clones were in vivo excised to give a population of pBluescript SK(-) phagemids. All steps to this point were performed in the ML Roose lab at the University of California, Riverside by X. Ye. Phagemids were plated, plasmid DNA purified, cDNA clones archived, and DNA sequences determined bi-directionally using an ABI3730 at the Arizona Genomics Institute, University of Arizona (Kim, Kudrna, Stum, Wissotski, Wing). Chromatogram files were downloaded to UC Riverside (Close), then processed at UC Riverside (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.

text: 10 - 30 cm shoots

text: E. coli TJC121

text: Citrus trifoliata

text: EcoRI

text: XhoI

text: Phloem

text: Poncirus trifoliata CTV-challenged cDNA library - AGI2

text: Phagemid pBluescript SK(-)

text: Lambda Uni-ZAP XR, excised phagemid


text: live

text: Generic

characteristic: NCBI submission model

text: Washington Navel

text: Parent Washington navel orange embryogenic callus was established from undeveloped ovules of ca. 10 mm diameter young fruits under open pollination on Murashige-Skoog medium at 25C with 16 h light in a tissue culture room. Embryogenic callus, globular and heart stage embryoids were pooled in approximately equal portions in RNAlater (Ambion), then RNA was extracted using TRIZOL Reagent (Invitrogen). Poly(A) RNA was purified from 500 microgram of total RNA using Qiagen Oligotex. A primary cDNA library was produced using a lambda ZAP XR cDNA Synthesis Kit (Stratagene). These steps were performed by Xinrong Ye (Roose lab, UC Riverside). One million pfu from the primary library were mass excised to produce a phagemid population by Raymond Fenton (Close lab, UC Riverside). Phagemids were plated, plasmid DNA purified, cDNA clones archived, and DNA sequences determined bi-directionally using an ABI3730 at the Arizona Genomics Institute, University of Arizona (Kim, Kudrna, Stum, Wissotski, Wing). Chromatogram files were downloaded by FTP to UC Riverside (by Close), then processed at UC Riverside (by Wanamaker, Close lab) 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.

text: Embryogenic and embryoid

text: E. coli TJC121

text: Citrus sinensis

text: EcoRI

text: XhoI

text: Callus

text: Parent Washington Navel Orange Callus cDNA Library UCRCS08-1

text: Phagemid pBluescript SK(-)

text: Lambda Uni-ZAP XR, excised phagemid


text: LIPM, CNRS/INRA

text: live

text: Generic

characteristic: NCBI submission package

text: Jemalong A17

text: The SSH-cDNA fragments were generated using the SSH-adaptor sequences ctaatacgactcactatagggctcgagcggccgcccgggcaggt and ctaatacgactcactatagggcagcgtggtcgcggccgaggt

text: 14 days old roots nitrogen starved for 4 days and 10-8M NodSm factor treated

text: E. coli ElectroTen-Blue electroporation-competent cells

text: Medicago truncatula

text: whole roots

text: MtSNF

text: pGEM-T


text: LIPM, CNRS/INRA

text: live

text: Generic

text: Jemalong A17

characteristic: description

text: 2 week old plants nitrogen starved for 4 days

text: Medicago truncatula

text: whole roots

text: pGEM-T


text: LIPM, CNRS/INRA

text: live

text: Generic

text: Jemalong A17

text: 14 days old roots nitrogen starved for three days

characteristic: lab-host

text: Medicago truncatula

text: whole roots

text: MtSCF

text: pGEM-T


text: LIPM, CNRS/INRA

text: live

text: Generic

text: Jemalong A17

text: 14-day-old roots, nitrogen-starved for three days

text: Medicago truncatula

characteristic: organism

text: whole roots

text: pGEM-T


text: LIPM, CNRS/INRA

text: live

text: Generic

text: Jemalong super nodulating mutant TR122 (allelic to Sunn)

text: 17 and 20 day old roots, 3 days and 6 days after inoculation by WT S. meliloti

characteristic: dev-stage

text: Medicago truncatula

text: whole roots

text: MtSTW

text: pGEM-T


text: LIPM, CNRS/INRA

text: live

text: Generic

text: 17 and 20 day old roots, 3 days and 6 days after inoculation by nodA- S.meliloti

text: Medicago truncatula

text: whole roots

characteristic: tissue-type

text: MtSTA

text: pGEM-T


text: LIPM, CNRS/INRA

text: live

text: Generic

text: Jemalong A17

text: Nodules 4 days and 10 days after Sinorhizobium meliloti inoculation (pooled)

text: Medicago truncatula

text: root nodules

characteristic: title

text: pGEM-T


text: Institute of Biochemical Plant Pathology, GSF

text: live

text: Generic

text: Nandu

text: 84 days

text: Triticum aestivum

text: flag leaves

text: Triticum aestivum flag leaves 84 days


text: RAU-University

text: live

text: Generic

text: cultured cells

text: Gossypium hirsutum

text: Cultured cotton cells


text: RAU-University

text: live

text: Generic

text: Gossypium hirsutum

text: leaf

text: Cotton leaf tissue


text: Auburn University

text: live

text: Generic

text: One year old C. unshui plants were grown for 5 weeks in a growth chamber with a 12 h light period at 400_mol m-2s-1 intensity. The temperature decline regime was as follows: 32_C day/21_C night (14 days); 27_C day/16_C night (7days); 24_C day/13_C night (7days) and 18_C day/7_C night (7days). RNA from the end of second week was used as an unacclimated control and RNA from the end of fifth week as treatment. mRNA Differential display was performed using six RNAimage kits and a total of 144 primer pairs according to the protocol enclosed (GenHunter, Nashiville, TN, U.S.A.).

text: E. coli

text: Citrus unshiu

text: Citrus unshiu cold acclimation with DDRT-PCR

text: pGEM-T Easy Vector


text: Stanford University Medical Center

text: live

text: Generic

text: Total RNA from e7.5 (80ug) embryos was used to generate cDNA using the Superscript plasmid system (Invitrogen, Carlsbad, CA). cDNA was cloned unidirectionally into pCS105. XL1Blue electrocompetent cells were transformed with ligated cDNAs. The resulting library has >500,000 clones and an average cDNA insert size of 1.7 kb.

text: Mus musculus

text: SalI

text: NotI

characteristic: strain

text: Plasmid


text: Stanford University Medical Center

text: live

text: Generic

text: Total RNA from e6.5 (80ug) embryos was used to generate cDNA using the Superscript plasmid system (Invitrogen, Carlsbad, CA). cDNA was cloned unidirectionally into pCS105. XL1Blue electrocompetent cells were transformed with ligated cDNAs. The resulting library has >500,000 clones and an average cDNA insert size of 1.7 kb.

text: Mus musculus

text: SalI

text: NotI

text: Plasmid

characteristic: vector


text: Urefv, Inra, INRA, Centre de Bordeaux

text: live

text: Generic

text: Fantasia

text: 30 days after bloom

text: Prunus persica

text: fruit mesocarp plus epidermis

text: Prunus persica fruit mesocarp plus epidermis 30 days after bloom


text: Urefv, Inra, INRA, Centre de Bordeaux

text: live

text: Generic

text: Fantasia

text: 80 days after bloom

text: Prunus persica

text: fruit mesocarp plus epidermis

text: Prunus persica fruit mesocarp plus epidermis 80 days after bloom


text: Chinese National Human Genome Center at Shanghai

text: live

text: Generic

text: Schistosoma japonicum long fragment cDNA

text: Schistosoma japonicum

text: SJL


text: National Cancer Institute / NIH

text: live

text: Generic

text: RNA obtained from pluripotent cell line derived from blastocyst inner cell mass (cell line HSF-6, NIH Registry designation UC06. Positive for OCT4 expression by rtPCR, positive for SSEA-3, SSEA-4, Tra-1-81, Tra-1-60 by immunofluorescence. Negative for SSEA-1 by immunofluorescence Passage 62. cDNA was primed using oligo-dT primer: 5'-pGACTAGTTCTAGATCGCGAGCGGCCGCCC(T)25-3' and cloned into the EcoRV/NotI sites of pExpress-1. Size-selection >1.25 kb resulted in an average insert size of 1.8 kb. This primary library is non-normalized (normalized primary library is NIH_MGC_281) and was constructed by Express Genomics (Frederick, MD). Note: this is a Mammalian Gene Collection library.

text: Blastocyst

characteristic: organ

text: Homo sapiens

text: EcoRV

text: NotI

text: plasmid


text: National Cancer Institute / NIH

characteristic: INSDC center name

text: live

text: Generic

text: RNA obtained from pluripotent cell line derived from blastocyst inner cell mass (cell line HSF-6, NIH Registry designation UC06. Positive for OCT4 expression by rtPCR, positive for SSEA-3, SSEA-4, Tra-1-81, Tra-1-60 by immunofluorescence. Negative for SSEA-1 by immunofluorescence Passage 62. cDNA was primed using oligo-dT primer: 5'-pGACTAGTTCTAGATCGCGAGCGGCCGCCC(T)25-3' and cloned into the EcoRV/NotI sites of pExpress-1. Size-selection >1.25 kb resulted in an average insert size of 2.0 kb. This primary library is normalized (non-normalized primary library is NIH_MGC_280) and was constructed by Express Genomics (Frederick, MD). Note: this is a Mammalian Gene Collection library.

text: DH10B TonA

text: Blastocyst

text: Homo sapiens

text: EcoRV

text: NotI

text: plasmid


text: National Cancer Institute / NIH

text: live

text: Generic

text: RNA obtained from pluripotent cell line derived from blastocyst inner cell mass (cell line HSF-1.14, NIH Registry designation UC01. Positive for OCT4 expression by rtPCR, positive for SSEA-3, SSEA-4, Tra-1-81, Tra-1-60 by immunofluorescence. Negative for SSEA-1 by immunofluorescence. Passage 35. This line is a subclone of the parental line; the parental line was subcloned to remove aneuploid cells). cDNA was primed using oligo-dT primer: 5'-pGACTAGTTCTAGATCGCGAGCGGCCGCCC(T)25-3' and cloned into the EcoRV/NotI sites of pExpress-1. Size-selection >1.25 kb resulted in an average insert size of 1.82 kb. This primary library is normalized (non-normalized primary library is NIH_MGC_278) and was constructed by Express Genomics (Frederick, MD). Note: this is a Mammalian Gene Collection library.

text: DH10B TonA

text: Blastocyst

text: Homo sapiens

text: EcoRV

text: NotI

text: plasmid


text: National Cancer Institute / NIH

text: live

text: Generic

text: RNA obtained from pluripotent cell line derived from blastocyst inner cell mass (cell line HSF-1.14, NIH Registry designation UC01. Positive for OCT4 expression by rtPCR, positive for SSEA-3, SSEA-4, Tra-1-81, Tra-1-60 by immunofluorescence. Negative for SSEA-1 by immunofluorescence. Passage 35. This line is a subclone of the parental line; the parental line was subcloned to remove aneuploid cells). cDNA was primed using oligo-dT primer: 5'-pGACTAGTTCTAGATCGCGAGCGGCCGCCC(T)25-3' and cloned into the EcoRV/NotI sites of pExpress-1. Size-selection >1.25 kb resulted in an average insert size of 1.9 kb. This primary library is non-normalized (normalized primary library is NIH_MGC_279) and was constructed by Express Genomics (Frederick, MD). Note: this is a Mammalian Gene Collection library.

text: DH10B TonA

text: Blastocyst

text: Homo sapiens

text: EcoRV

text: NotI

text: plasmid


text: live

text: Generic

text: The library was constructed according Bonaldo, Lennon and Soares, Genome Research, 6:791-806, 1996. Denatured RNA was size fractionated on a 1% agarose gel.First strand cDNA synthesis was primed with oligo-dT primer containing a Not I site .Double strand cDNA was size selected according to mRNA size fraction, ligated with EcoR I adaptor, digested with NotI and then cloned directionally into pYX-Asc vector. The library tag sequences located between the Not I site and the polyA tail are CTGCGTCCTC and AGAGGAGACG. This library was created for the University Iowa Brain Anatomy Project (BMAP): 'Gene Discovery in the Developing Mouse Nervous System', supported by National Institute of Mental Health (NIMH), Hemin Chin, Ph.D., program coordinator.

text: embryos 15,16,17,18 dpc

text: DH10B (T1 phage resistant)

text: Eye

text: Mus musculus

text: EcoR I

text: Not I

text: whole eye

text: Plasmid

text: pYX-Asc


text: Washington University School of Medicine

text: live

text: Generic

text: Testis cDNA Library, Drosophila yakuba, strain Tai18E2. 1) Testes were dissected from 100 flies in RNA Later (Ambion); stored tissue in RNA Later. 2) Total RNA isolated from testes using Ambion MirVana Kit. 3) Invitrogen GeneRacer Kit used to generate cDNAs derived from processed transcripts (inserts have GeneRacer RNA Oligo ligated to the 5' ends and GeneRacer Oligo dT Primer ligated to the 3' ends). 4) cDNA was amplified for 5 cycles using Roche Expand High Fidelity PCR System and cleaned up using Qiagen QIAquick PCR Purification Kit. 5) Purified cDNAs incubated with dATP, buffer and Promega Taq DNA Polymerase @ 72C for 15 minutes. 6) cDNA ligated into Invitrogen PCR-4 TOPO plasmid vector. Approximate titer: Transformation of 2ul of ligation product into Invitrogen TOP10 cells yielded roughly 1000 colonies. 5' ends can be sequenced using GeneRacer 5' nested primer. GeneRacer RNA Oligo Sequence: 5'-CGACUGGAGCACGAGGACACUGACAUGGACUGAAGGAGUAGAAA-3' ; GeneRacer 5' Nested Primer Sequence: 5'-GGACACTGACATGGACTGAAGGAGTA-3' ; GeneRacer Oligo dT Primer Sequence: 5'-GCTGTCAACGATACGCTACGTAACGGCATGACAGTG(T)24-3' ; GeneRacer 3' Nested Primer Sequence: 5'-CGCTACGTAACGGCATGACAGTG-3'

text: TOP10 cells (Invitrogen)

text: Drosophila yakuba

text: D yakuba testes cDNA

text: plasmid

text: PCR-4 TOPO (Invitrogen)


text: Northeast Forestry University

text: live

text: Generic

text: Tamarix androssowii leaves were treated with NaHCO3 for 48 hours before cDNA library construction.

text: E. coli strain SOLR

text: Tamarix androssowii

text: EcoRI

text: XhoI

text: leaf

text: cDNA library of Tamarix androssowii

text: Plasmid

text: pBluescript SK(-)


text: RMIT University

text: live

text: Generic

characteristic: cultivar

text: Prepared with Clontech SMART PCR cDNA Synthesis kit, pGEM-T Easy vector, E. coli JM109, QIAGEN mini-prep kit, Big-Dye sequencing

text: 2 weeks and 24-48 hours post-inoculation with Ascochyta rabiei

text: Cicer arietinum

text: Stem and leaf

text: Chickpea (ICC3996) post-inoculation EST library

text: pGEM-T Easy


characteristic: External Id

text: College of Agronomy and Biotechnology, China Agricultural University

text: live

text: Generic

text: Gossypium hirsutum

text: cotton fiber

text: cotton fiber


text: Northeast Forestry University

characteristic: INSDC first public

text: live

text: Generic

text: Tamarix ramosissima

text: leaf

text: Tamarix ramosissim Ledeb. suppression subtractive hybridization cDNA library


text: Children's Memorial Research Center

characteristic: INSDC last update

text: live

text: Generic

text: UI-R-DZ1 is a normalized cDNA library containing the following tissue(s): Swarm Rat Chondrosarcoma. The library was constructed according to Bonaldo, Lennon and Soares, Genome Research, 6:791-806, 1996. First strand cDNA synthesis was primed with an oligo-dT primer containing a Not I site. Double stranded cDNA was ligated to an EcoR I adaptor, digested with Not I, and cloned directionally into pT7T3-Pac vector. The oligonucleotide used to prime the synthesis of first-strand cDNA contains a library tag sequence that is located between the Not I site and the (dT)18 tail. The sequence tag for this library is CATTCTTGTA. The Rat cartilaginous tumor tissue was provided by Dr Jeff Stevens at the University of Iowa.

text: 37 days

text: DH10B (Life Technologies)

text: Spine

text: Rattus norvegicus

text: Not I

text: Eco RI

text: Sprague-Dawley

text: Chondrosarcoma

text: Phagemid


text: University of Western Ontario

text: live

characteristic: INSDC status

text: Generic

text: Apis mellifera

text: Honey bee EST


text: Royal College of Surgeons in Ireland

text: live

text: Generic

characteristic: NCBI submission model

text: Mus musculus

text: SalI

text: NotI

text: Plasmid

text: pQE-30NASTattB (GenBank-Accession-Nummer: AY386205)


text: Institute of Biotechnology of Leon (Inbiotec)

text: live

text: Generic

characteristic: NCBI submission package

text: cDNA molecules expressed uniquely or preferentially in glucose-containing medium obtained by suppression subtractive hybridization using the Clontech PCR-SelectTM cDNA subtraction kit. The PCR-amplified cDNA fragments were then ligated to a EcoRV-digested pZeRO-2.1 vector and transformed into E. coli TOP-10

text: Mycelium grown for 24 h in glucose-containing CP medium

text: Penicillium chrysogenum

text: NRRL1951 (wild-type)

text: cDNA expressed in glucose-containing medium

text: Plasmid


text: live

text: Generic

text: cDNA molecules expressed uniquely or preferentially in lactose-containing medium obtained by suppression subtractive hybridization using the Clontech PCR-SelectTM cDNA subtraction kit. The PCR-amplified cDNA fragments were then ligated to a EcoRV-digested pZeRO-2.1 vector and transformed into E. coli TOP-10

characteristic: description

text: Mycelium grown for 24 h in lactose-containing CP medium

text: Penicillium chrysogenum

text: NRRL1951 (wild-type)

text: cDNA expressed in lactose-containing medium

text: Plasmid


text: Washington University School of Medicine

text: live

text: Generic

text: GoldRush

text: Total RNA was extracted from freeze dried leaf tissue Stage II, using the method described by Wang and Vodkin (Plant Molecular Biology Reporter 12:132-145, 1994). Poly(A)+ mRNA was isolated from total RNA using the PolyATtract mRNA Isolation system III (Promega). The library was prepared using the Stratagene pBluescript II XR cDNA library construction kit. Complementary DNA was synthesized from mRNA using a Poly (dT) sequence with a XhoI restriction site. EcoRI adapters were ligated to the blunt ended cDNA fragments followed by restriction with XhoI. The cDNA insert is protected from XhoI digestion via methylation during first strand cDNA synthesis. The cDNA fragments were directionally cloned into the EcoRI-XhoI restriction site of the pBluescript vector. The ligated cDNA fragments were transformed into E. coli ElectroMax DH10B host cells. Transformation efficiency: 3.6E+07 Average insert Size by PCR: 1053 bp

text: DH10B ampicillin resistant

characteristic: lab-host

text: Malus domestica

text: Xho I

text: Eco RI

text: Leaf

text: Phagemid (ampicillin resistant)

text: pBluescript II SK (+)


text: Tokyo University of Marine Science and Technology

text: live

text: Generic

text: Adult

text: Paralichthys olivaceus

characteristic: organism

text: Liver

text: Japanese flounder liver cDNA library

text: pSPORT 1


text: The Institute of Tropical Medicine

text: live

text: Generic

text: adult worms

characteristic: dev-stage

text: Angiostrongylus cantonensis

text: Angiostrongylus cantonensis adult worms cDNA clone


text: The Institute of Tropical Medicine

text: live

text: Generic

text: 3 weeks infection

text: Angiostrongylus cantonensis

text: Angiostrongylus cantonensis 3 weeks infection cDNA clone


text: INRA

text: live

text: Generic

text: Cabernet-Sauvignon

text: Vitis vinifera

text: Vitis vinifera Cabernet-Sauvignon root


text: INRA

text: live

text: Generic

text: Cabernet-Sauvignon

text: Vitis vinifera

text: Vitis vinifera Cabernet-Sauvignon fruit


text: INRA

text: live

text: Generic

text: Cabernet-Sauvignon

text: Vitis vinifera

text: Vitis vinifera Cabernet-Sauvignon leaf


text: UMR 7100 - IFR 85 - ESBS -ULP

text: live

text: Generic

text: The subtractive hybridization protocol from BD Biosciences Clontech (Palo Alto, CA) has been used to generate this specific cDNA library. Tester cDNA was prepared from mycelium grown in minimal medium M3 containing hop cell walls as sole carbon source and driver cDNA was prepared from mycelium grown in minimal M3 containing glucose. The resulting cDNAs were cloned into a ddTTP tailed pUC19 vector.

text: E.coli TOP10 F'

text: Fusarium graminearum

text: HindIII

text: DIFCW SSH cDNA

text: Plasmid


text: INRA Rennes

text: live

text: Generic

text: Sample name: ID0ACC ; Plant growth place: INRA Rennes, UMR BiO3P, 35327, 35653 Le Rheu Cedex France ; Soil conditions: Soil ; Sowing date: 01/10/2003 ; Harvesting date: 17/10/2003 ; Description: aphids inoculated on one-week old Vicia faba germinations under non sterile conditions experimental condition: long photoperiod (16-hr light/8-hr dark at 18 degc)

text: larvae L3 (parthenogenetic females)

text: Acyrthosiphon pisum

text: EcoRI

text: XhoI

text: head

characteristic: tissue-type

text: pBS-SKminus


text: INRA Rennes

text: live

text: Generic

text: Sample name: ID0ADD ; Plant growth place: INRA Rennes, UMR BiO3P, 35327, 35653 Le Rheu Cedex France ; Soil conditions: Soil ; Sowing date: 15/06/2004 ; Harvesting date: 15/06/2004 ; Description: Aphids inoculated on one-week old Vicia faba under non-sterile conditions. A. pisum YR2 is holocyclic, i.e. able to change its reproductive mode under short photoperiods (sexual) versus long photoperiods (clonal). experimental condition: long photoperiod (16-hr light/8-hr dark at 18 degc)

text: larvae L3

text: Acyrthosiphon pisum

text: EcoRI

text: XhoI

text: embryos

characteristic: title

text: pBS-SKminus


text: INRA Rennes

text: live

text: Generic

text: Sample name: ID0AEE ; Plant growth place: INRA Rennes, UMR BiO3P, 35327, 35653 Le Rheu Cedex France ; Soil conditions: Soil ; Sowing date: 15/04/2004 ; Harvesting date: 15/04/2004 ; Description: Aphids inoculated on one-week old Vicia faba under non-sterile conditions. A. pisum YR2 is holocyclic, i.e. able to change its reproductive mode under short photoperiods (sexual) versus long photoperiods (clonal). experimental condition: long photoperiod (16-hr light/8-hr dark at 18 degc)

text: Acyrthosiphon pisum

text: EcoRI

text: XhoI

text: antennae

text: pBS-SKminus


text: School of Agricultural Biotechnology, Seoul National University

text: live

text: Generic

text: The libraries were synthesized using the ZAP(r)-cDNA synthesis method (Stratagene). cDNA was prepared, size-fractionated and inserted into the Uni-ZAP XR vector using XhoI linker-primer and EcoRI adaptor. After in vivo excision with E. coli strain SOLR, cDNA libraries from testis contain each insert size range of 0.5~3kb (n=20), and 0.5~3.5kb (n=20).

text: 6.5 day embryo

text: Gallus gallus

text: XhoI

text: EcoRI

text: Male+Female

characteristic: sex

text: White Leghorn

characteristic: strain

text: Gonad

text: Embryonic gonad cDNA Library

text: phage

text: Uni-ZAP XR vector


text: Dept Biological Sciences, National University of Singapore

text: live

text: Generic

text: Hippocampus comes

text: Male

text: Seahorse brood pouch Uni-ZAP XR library

text: Uni-ZAP XR

characteristic: vector


text: University of New Mexico

characteristic: INSDC center name

text: live

text: Generic

text: M-line strain B. glabrata were infected with Echinostoma paraensei. At t=48 hours, mRNA was harvested from infected and uninfected snails. cDNA was generated using oligo-dT primers. The B. glabrata M line (E. paraensei-exposed) cDNA was subtracted against M-line cDNA using the PCR-Select Suppressive Subtraction Hybridization kit (SSH) (Clontech, Palo Alto, CA). The resulting library was cloned into pCR2.1 (Invitrogen, Carlsbad, CA) and sequenced using plasmid-specific primers (M13 rev and/or M13 -20). Primers were removed from the sequences obtained. cames = C Adema M-line Echinostoma (paraensei) SSH-EST.

text: adult

text: Biomphalaria glabrata

text: hermaphrodite

text: M line

text: whole body

text: B. glabrata M line (E. paraensei exposed) - M line (control) SSH library

text: Plasmid


text: Union Hospital of Fujian Medical University

text: live

text: Generic

text: leukaemic

characteristic: cell type

text: adolescent

text: Homo sapiens

text: male

text: bone marrow

text: Homo sapiens bone marrow adolescent


characteristic: External Id

text: live

text: Generic

text: Leukemia cells

text: Adolescent

text: Homo sapiens

text: HindIII

text: NotI

text: male

text: Bone marrow

text: Human adolescent leukemia cells HindIII-NotI

text: Plasmid


characteristic: INSDC first public

text: live

text: Generic

text: Leukemia cells

text: Adolescent

text: Homo sapiens

text: EcoRI

text: EcoRI

text: male

text: Bone marrow

text: Human adolescent leukemia cells EcoRI

text: Plasmid


text: University of California

characteristic: INSDC last update

text: live

text: Generic

text: Ruby

characteristic: cultivar

text: Two trees with open-pollinated flowers in a mixed planting at UC Riverside Agricultural Operations were the source of tissue. Flower buds were collected by Federici (Roose lab) from January -March 2003 in four groups: 1) no white petals, 2) white petals visible, bud less than 0.5 cm, 3) bud more than 0.5 cm but not open, 4) newly opened flowers. Tissues were snap frozen and then stored at -80C until further processing. Fenton (Close lab) purified RNA by the phenol method described in J. Japanese Soc. Hort. Sci. 1996. 64 (4): 809-814, purified poly(A) mRNA using a PolyATtrack mRNA Isolation System IV (Promega), produced a primary cDNA library using a lambda ZAP XR cDNA Synthesis Kit (Stratagene), then mass-excised one million pfu from the primary library to produce a phagemid population. Phagemids were plated, plasmid DNA purified, cDNA clones archived, and DNA sequences determined bi-directionally using an ABI3730 at the Genomics Core Instrumentation Facility, University of California, Riverside (Choi, Kingan). Chromatogram files were downloaded by FTP by Close, then processed by Wanamaker (Close lab) 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.

text: 20 year old trees

text: E. coli TJC121

text: Citrus sinensis

text: EcoRI

text: XhoI

text: Flower

text: Ruby Orange Developing Flower cDNA Library UCRCS04-UCR

text: Phagemid pBluescript SK(-)

text: Lambda Uni-ZAP XR, excised phagemid


text: Animal and Natural Resources Institute

text: live

characteristic: INSDC status

text: Generic

characteristic: NCBI submission model

text: Standard dT, Non-normalized library of pooled RNA isolated from whole brain, ultimobranchial gland, parathyroid gland, cecal tonsil, and primordial germ cells multiple

text: Gallus gallus

text: NotI

text: EcoRI

text: Leghorn and Broiler

text: BARC_4GAL chicken mixed tissue

text: plasmid

text: pBluescript, SK+, Stratagene


text: University of California

text: live

text: Generic

characteristic: NCBI submission package

text: Phytophthora infestans

text: mycelium, non-sporulating growth

text: Plasmid


text: University of California

text: live

text: Generic

text: Phytophthora infestans

text: mycelium, sporulating growth

text: Plasmid


text: University of California

text: live

text: Generic

text: Phytophthora infestans

text: mycelium, nitrogen starvation

text: Plasmid


text: University of California

text: live

text: Generic

text: Phytophthora infestans

text: mycelium, carbon starvation

text: Plasmid


text: University of California

text: live

text: Generic

text: Phytophthora infestans

text: sporangia, cleaving

text: Plasmid


text: University of California

text: live

text: Generic

text: Phytophthora infestans

text: sporangia, germinating

text: Plasmid


text: University of California

text: live

text: Generic

text: Phytophthora infestans

text: cysts, germinating

text: Plasmid


text: University of California

text: live

text: Generic

text: Phytophthora infestans

text: infected tomato, outside of lesion 3 dpi

text: Plasmid


text: University of California

text: live

text: Generic

text: Phytophthora infestans

text: infected tomato, center of lesion 3 dpi

text: Plasmid


text: University of California

text: live

text: Generic

text: Phytophthora infestans

text: sporangia, purified

text: Plasmid


text: University of California

text: live

text: Generic

text: Phytophthora infestans

text: mycelium, heat treated

text: Plasmid


text: University of California

text: live

text: Generic

text: Phytophthora infestans

text: mycelium, H2O2-treated

text: Plasmid


text: University of California

text: live

text: Generic

text: Phytophthora infestans

text: A1 and A2

text: 88069 and 618

text: mating of 88069 (A1) and 618 (A2)

text: Plasmid


text: University of California

text: live

text: Generic

text: Phytophthora infestans

text: infected potato, center of lesion 6 dpi

text: Plasmid


text: University of California

text: live

text: Generic

text: Phytophthora infestans

text: infected potato, outside of lesion 6 dpi

text: Plasmid


text: University of California

text: live

text: Generic

text: Phytophthora infestans

text: mycelium, subtracted infection mimic

text: Plasmid


text: University of California

text: live

text: Generic

text: Phytophthora infestans

text: zoospores, purified

text: Plasmid


text: University of California

text: live

text: Generic

text: Phytophthora infestans

text: mycelium, infection mimic

text: Plasmid


text: University of California

text: live

text: Generic

text: Phytophthora infestans

text: mycelium, starved in water

text: Plasmid


text: University of California

text: live

text: Generic

text: Phytophthora infestans

text: mycelium, Plich medium

text: Plasmid


text: Cornell University

text: live

text: Generic

text: Signal sequence trap library of the tomato late blight interactome

characteristic: description

text: Phytophthora infestans

text: PiE


text: Glasgow Caledonian University

text: live

text: Generic

text: Subtracted cDNA library from liver of immature EE2-exposed male plaice

text: Immature

text: Liver

characteristic: organ

text: Pleuronectes platessa

characteristic: organism

text: Male

text: Plaice cDNA library


text: Queens University of Belfast, Grosvenor Road, Belfast BT12 6BJ, UK

text: live

text: Generic

text: Breast metastatic cell line

characteristic: cell-line

text: Subtracted cDNA Library

text: Rattus norvegicus

text: Breast metastatic cell line


text: live

text: Generic

text: Breast cancer metastatic cell line

text: Subtracted cDNA Library

text: Rattus norvegicus

text: Breast cancer metastatic cell line

characteristic: title


text: University of Hamburg, Biocenter Klein Flottbek

text: live

text: Generic

text: 2-cell proembryo

text: Florida

text: 24 hours after fertilization

characteristic: dev-stage

text: Triticum aestivum


text: live

text: Generic

text: egg cell

text: Florida

text: unfertilized

text: Triticum aestivum

text: EC


text: West China Hospital, Sichuan University

text: live

text: Generic

text: oocytes

text: Xenopus laevis

text: EcoRI

text: XbaI

text: Xenopus laevis Lambda TriplEx cDNA Express Library

text: phagemid

text: pTriplEx


text: University of Missouri-Columbia

text: live

text: Generic

text: Funding: The production of ESTs submitted in this project was funded by USDA Grant MRI-2002-03476 entitled 'Bovine ESTs: Focus on Female Reproduction' to RS Prather(Primary Investigator), E Antoniou, HA Garverick, JA Green, MC Lucy, RM Roberts, MF Smith and RS Youngquist. Genetic Source: Heifers for the project were purchased from Circle A Ranch, Iberia, MO (http://www.circlearanch.com/home.html). These heifers, while not registered have known Angus pedigrees going back at least 4 generations. Samples collected: The samples consisted of the following: germinal vesicle-stage oocytes; in vitro derived embryos (2-cell, morula, blastocyst and nuclear transfer blastocyst); in vivo blastocysts and conceptuses (days 8, 14, 16 and 18); corpora lutea (days 3, 5, 8, 14, 16, 18 and 35); ovarian follicles (days 0, non-recruited, recruited, early selected and preovulatory); oviduct (days 0, 3 and 5); endometrium (days 5, 8, 14, 16, 18 and 35); and placenta/embryo from day 35 conceptuses. Expanded descriptions of how the tissues were collected can be found at the following URL: http://genome.rnet.missouri.edu/Bovine/Methods.html. Library Construction (PCR Protocol): The amount of mRNA that was recovered from oocytes and embryos was quite limiting and was not sufficient for library production with the standard protocol. Therefore, PCR-based protocol was utilized for producing libraries from sources in which the amount of extracted mRNA was small (oocytes and embryos). Poly-A RNA was isolated by using the MicroPoly(A) Pure kit from Ambion (cat. # 1918). The mRNA was reverse transcribed with a NotI-tag-dT18 oligonucleotide and a SMART oligonucleotide (Clontech) modified to contain a SalI site to generate full-length cDNA with a sequence complementary to the SMART oligonucleotide. Sequences within the SMART and dT oligonucleotides were used as primers to amplify the cDNAs by PCR with pfu turbo polymerase (Stratagene). The resulting PCR products were purified, digested with NotI and SalI and size fractionated by using Chroma Spin-1000 columns (Clontech). Purified cDNA from each PCR reaction was quantitated and mixed on an equimolar basis for ligation into the pCMV- SPORT6 vector. Preliminary Library Characterization: Randomly chosen clones from each library were analyzed by restriction digestion to determine average insert size (96 clones) and by sequencing (~4 96-well plates) to confirm library quality [e.g. the presence of short polyA+ tails, genomic DNA contamination (must be <1%), ribosomal RNA clones (must be <1%), etc.] and to provide a sequence database representing the predominant clones in each library. The clones were sequenced at the University of Missouri-Columbia DNA Core Facility. After production of the libraries, equal numbers of recombinants from each library were pooled to produce a single mixed library (mega-library) for more extensive sequencing. Bioinformatics work was performed by GK Springer's bioinformatics group in Computer Science at the University of Missouri-Columbia. Clone Requests: Requests for clones should be made to the Director of the University of Missouri DNA Core facility at: bovine@rnet.missouri.edu. Citations: Bonaldo MF, Lennon G, Soares MB,. Normalization and Subtraction: Two approaches to facilitate gene discovery. Genome Res, 1996; 6:791-806. Jiang H, Bivens NJ, Ries JE, Whitworth KM, Green JA, Forrester LJ, Springer GK, Didion BA, Mathialagan N, Prather RS, Lucy MC (2001) Constructing cDNA libraries with fewer clones that contain long poly(dA) tails. Biotechniques 31:38-42. Soares MB, MF Banaldo, P Jelene, L Su, L Lawton, A Efstrantiadis. 1994. Construction and characterization of a normalized cDNA library. Proc Natl Acad Sci, 91:9228-9232.

text: In vivo derived blastocysts Day 8

text: Bos taurus

text: bconb

text: Cloning


text: University of Missouri-Columbia

text: live

text: Generic

text: Funding: The production of ESTs submitted in this project was funded by USDA Grant MRI-2002-03476 entitled 'Bovine ESTs: Focus on Female Reproduction' to RS Prather(Primary Investigator), E Antoniou, HA Garverick, JA Green, MC Lucy, RM Roberts, MF Smith and RS Youngquist. Genetic Source: Heifers for the project were purchased from Circle A Ranch, Iberia, MO (http://www.circlearanch.com/home.html). These heifers, while not registered have known Angus pedigrees going back at least 4 generations. Samples collected: The samples consisted of the following: germinal vesicle-stage oocytes; in vitro derived embryos (2-cell, morula, blastocyst and nuclear transfer blastocyst); in vivo blastocysts and conceptuses (days 8, 14, 16 and 18); corpora lutea (days 3, 5, 8, 14, 16, 18 and 35); ovarian follicles (days 0, non-recruited, recruited, early selected and preovulatory); oviduct (days 0, 3 and 5); endometrium (days 5, 8, 14, 16, 18 and 35); and placenta/embryo from day 35 conceptuses. Expanded descriptions of how the tissues were collected can be found at the following URL: http://genome.rnet.missouri.edu/Bovine/Methods.html. Library construction (Standard Protocol): All procedures have been described in detail elsewhere (Soares et al., 1994; Bonaldo et al., 1996; Jiang et al., 2001). Total cellular RNA from each sample was isolated by using STAT-60 reagent (Tel-Test, Friendswood, TX) and the poly(A)+ RNA was obtained by two rounds of purification with the Oligotex mRNA isolation kit (Qiagen) according to the manufacturer's instructions. The libraries were constructed essentially as described by the manufacturer's instructions provided with the SuperScript Plasmid System (Invitrogen, cat. no. 18248-013). Briefly, 1mcg of poly(A)+ RNA was annealed at c37 degrees with 10mcg of NotI-tag-dT18 oligonucleotide (GCTGCTCGCGGCCGC-tag-T18) and reverse transcribed at c37 degrees with SuperScript II (Invitrogen) reverse transcriptase (Jiang et al., 2001). The 'tag' represents a tissue/stage-specific ten-base sequence identifier (http://genome.uiowa.edu/pubsoft/software.html) present in the oligonucleotide used to prime first-strand synthesis. Second strand synthesis was performed with T4 DNA polymerase in the presence of DNA ligase and RNase H. After second strand synthesis, the double-stranded cDNAs were ligated to SalI adapters (Invitrogen-Life Technologies) and digested with NotI. The cDNAs were size selected by passage through cDNA size fractionation columns (Invitrogen-Life technologies). The cDNAs derived from each developmental stage of a particular tissue were mixed on an equimolar basis and ligated directionally into the NotI and SalI sites of the pCMV-SPORT6 vector (Invitrogen). After ligation of the inserts, the plasmids were electroporated into DH10B bacteria. Preliminary Library Characterization: Randomly chosen clones from each library were analyzed by restriction digestion to determine average insert size (96 clones) and by sequencing (~4 96-well plates) to confirm library quality [e.g. the presence of short polyA+ tails, genomic DNA contamination (must be <1%), ribosomal RNA clones (must be <1%), etc.] and to provide a sequence database representing the predominant clones in each library. The clones were sequenced at the University of Missouri-Columbia DNA Core Facility. After production of the libraries, equal numbers of recombinants from each library were pooled to produce a single mixed library (mega-library) for more extensive sequencing. Bioinformatics work was performed by GK Springer's bioinformatics group in Computer Science at the University of Missouri-Columbia. Clone Requests: Requests for clones should be made to the Director of the University of Missouri DNA Core facility at: bovine@rnet.missouri.edu. Citations: Bonaldo MF, Lennon G, Soares MB,. Normalization and Subtraction: Two approaches to facilitate gene discovery. Genome Res, 1996; 6:791-806. Jiang H, Bivens NJ, Ries JE, Whitworth KM, Green JA, Forrester LJ, Springer GK, Didion BA, Mathialagan N, Prather RS, Lucy MC (2001) Constructing cDNA libraries with fewer clones that contain long poly(dA) tails. Biotechniques 31:38-42. Soares MB, MF Banaldo, P Jelene, L Su, L Lawton, A Efstrantiadis. 1994. Construction and characterization of a normalized cDNA library. Proc Natl Acad Sci, 91:9228-9232.

text: Conceptus-Corpus Luteum(mixed)

text: Bos taurus

text: bmix

text: Cloning


text: University of Missouri-Columbia

text: live

text: Generic

text: Funding: The production of ESTs submitted in this project was funded by USDA Grant MRI-2002-03476 entitled 'Bovine ESTs: Focus on Female Reproduction' to RS Prather(Primary Investigator), E Antoniou, HA Garverick, JA Green, MC Lucy, RM Roberts, MF Smith and RS Youngquist. Genetic Source: Heifers for the project were purchased from Circle A Ranch, Iberia, MO (http://www.circlearanch.com/home.html). These heifers, while not registered have known Angus pedigrees going back at least 4 generations. Samples collected: The samples consisted of the following: germinal vesicle-stage oocytes; in vitro derived embryos (2-cell, morula, blastocyst and nuclear transfer blastocyst); in vivo blastocysts and conceptuses (days 8, 14, 16 and 18); corpora lutea (days 3, 5, 8, 14, 16, 18 and 35); ovarian follicles (days 0, non-recruited, recruited, early selected and preovulatory); oviduct (days 0, 3 and 5); endometrium (days 5, 8, 14, 16, 18 and 35); and placenta/embryo from day 35 conceptuses. Expanded descriptions of how the tissues were collected can be found at the following URL: http://genome.rnet.missouri.edu/Bovine/Methods.html. Library construction (Standard Protocol): All procedures have been described in detail elsewhere (Soares et al., 1994; Bonaldo et al., 1996; Jiang et al., 2001). Total cellular RNA from each sample was isolated by using STAT-60 reagent (Tel-Test, Friendswood, TX) and the poly(A)+ RNA was obtained by two rounds of purification with the Oligotex mRNA isolation kit (Qiagen) according to the manufacturer's instructions. The libraries were constructed essentially as described by the manufacturer's instructions provided with the SuperScript Plasmid System (Invitrogen, cat. no. 18248-013). Briefly, 1mcg of poly(A)+ RNA was annealed at c37 degrees with 10mcg of NotI-tag-dT18 oligonucleotide (GCTGCTCGCGGCCGC-tag-T18) and reverse transcribed at c37 degrees with SuperScript II (Invitrogen) reverse transcriptase (Jiang et al., 2001). The 'tag' represents a tissue/stage-specific ten-base sequence identifier (http://genome.uiowa.edu/pubsoft/software.html) present in the oligonucleotide used to prime first-strand synthesis. Second strand synthesis was performed with T4 DNA polymerase in the presence of DNA ligase and RNase H. After second strand synthesis, the double-stranded cDNAs were ligated to SalI adapters (Invitrogen-Life Technologies) and digested with NotI. The cDNAs were size selected by passage through cDNA size fractionation columns (Invitrogen-Life technologies). The cDNAs derived from each developmental stage of a particular tissue were mixed on an equimolar basis and ligated directionally into the NotI and SalI sites of the pCMV-SPORT6 vector (Invitrogen). After ligation of the inserts, the plasmids were electroporated into DH10B bacteria. Library Construction (PCR Protocol): The amount of mRNA that was recovered from oocytes and embryos was quite limiting and was not sufficient for library production with the standard protocol. Therefore, PCR-based protocol was utilized for producing libraries from sources in which the amount of extracted mRNA was small (oocytes and embryos). Poly-A RNA was isolated by using the MicroPoly(A) Pure kit from Ambion (cat. # 1918). The mRNA was reverse transcribed with a NotI-tag-dT18 oligonucleotide and a SMART oligonucleotide (Clontech) modified to contain a SalI site to generate full-length cDNA with a sequence complementary to the SMART oligonucleotide. Sequences within the SMART and dT oligonucleotides were used as primers to amplify the cDNAs by PCR with pfu turbo polymerase (Stratagene). The resulting PCR products were purified, digested with NotI and SalI and size fractionated by using Chroma Spin-1000 columns (Clontech). Purified cDNA from each PCR reaction was quantitated and mixed on an equimolar basis for ligation into the pCMV- SPORT6 vector. Preliminary Library Characterization: Randomly chosen clones from each library were analyzed by restriction digestion to determine average insert size (96 clones) and by sequencing (~4 96-well plates) to confirm library quality [e.g. the presence of short polyA+ tails, genomic DNA contamination (must be <1%), ribosomal RNA clones (must be <1%), etc.] and to provide a sequence database representing the predominant clones in each library. The clones were sequenced at the University of Missouri-Columbia DNA Core Facility. After production of the libraries, equal numbers of recombinants from each library were pooled to produce a single mixed library (mega-library) for more extensive sequencing. Bioinformatics work was performed by GK Springer's bioinformatics group in Computer Science at the University of Missouri-Columbia. Clone Requests: Requests for clones should be made to the Director of the University of Missouri DNA Core facility at: bovine@rnet.missouri.edu. Bonaldo MF, Lennon G, Soares MB,. Normalization and Subtraction: Two approaches to facilitate gene discovery. Genome Res, 1996; 6:791-806. Jiang H, Bivens NJ, Ries JE, Whitworth KM, Green JA, Forrester LJ, Springer GK, Didion BA, Mathialagan N, Prather RS, Lucy MC (2001) Constructing cDNA libraries with fewer clones that contain long poly(dA) tails. Biotechniques 31:38-42. Soares MB, MF Banaldo, P Jelene, L Su, L Lawton, A Efstrantiadis. 1994. Construction and characterization of a normalized cDNA library. Proc Natl Acad Sci, 91:9228-9232.

text: Ovarian Follicle

text: Bos taurus

text: bof

text: Cloning


text: Forestry and Forest Products Research Institute

text: live

text: Generic

text: cDNA-RDA

text: primordium

text: Lentinula edodes

characteristic: strain

text: Lentinula edodes A567 primordium


text: live

text: Generic

text: cDNA-RDA

text: mature fruiting body

text: Lentinula edodes

text: Lentinula edodes A567 mature fruiting body


text: Miyazaki University

text: live

text: Generic

text: Cyprinus carpio

text: head kidney

characteristic: tissue-type

text: Cyprinus carpio head kidney


text: Soybean Research Institute

characteristic: INSDC center name

text: live

text: Generic

text: Dongnong 8143

text: A SMV-resistant variety, Dongnong 8143 was used to construct the subtractive library by SSH after inoculating with SMV Strain No.1.

text: 0-10 days after inoculating by SMV No.1 when soybean is in VE stage seedling

text: Leaf

text: Glycine max

text: hermaphrodite

characteristic: sex

text: Soybean Disease-resistance Subtracted Library


characteristic: External Id

text: National Cancer Institute / NIH

text: live

text: Generic

text: human embryonic stem cells

characteristic: cell type

text: RNA obtained from human embryonic stem cells isolated from the inner cell mass of blastocyst stage embryos and differentiated to an early endodermal cell type. Cell line id and NIH Registry designation is BGO1. Positive for GATA4, MixL1, Msx1, HNF4alpha expression; negative for AFP expression. Passage number 40. cDNA primed using oligo-dT primer: 5'-pGACTAGTTCTAGATCGCGAGCGGCCGCCC(T)25-3' and cloned into the EcoRV/NotI sites of pExpress-1. This primary library is non-normalized (normalized primary library is NIH_MGC_259). It was constructed by Express Genomics (Frederick, MD). Sequence ends have been trimmed to exclude vector and regions below Phred quality 16. Three-prime sequences are presented as their reverse complement and have been trimmed to exclude polyA. Note: this is a Mammalian Gene Collection library.

text: DH10B-T1 phage-resistant E. coli

characteristic: lab-host

text: Homo sapiens

text: NotI

text: EcoRV

text: male

text: human embryonic stem cells differentiated to an early endodermal cell type

text: Plasmid

characteristic: vector


text: Tianjin central hospital of obstetrics and gynaecology

characteristic: INSDC first public

characteristic: INSDC last update

text: live

characteristic: INSDC status

text: Generic

text: The method of mRNA differential display was applied to find differential expressed cDNA bands in placenta between tne preeclampsia patient and normal gestation woman.

text: Homo sapiens

text: placenta

text: cDNA Clone of a Human Placenta


text: University of California

text: live

text: Generic

characteristic: NCBI submission model

characteristic: NCBI submission package

text: Parent Washington Navel

characteristic: cultivar

text: Trees were grown in the field at University of California, Riverside using standard horticultural practices. Young fruits were placed in a cage with thrips (Scirtothrips citri). Infestations were conducted by Watkins (Morse lab) and flavedo collected by Federici (Roose lab) from May-June 2003. The thrips were collected from naturally infested Rhus plants by sucking into a tube. A flexible hose was attached to a tube that extended into a covered vial, and another tube stuck out of the vial at a right angle. The bent tube was held above the thrips, sucking on the flexible tube created a vacuum, pulling the thrips into the vial. Thrips were knocked off the Rhus plant onto a manila folder, then only second instars were captured. This was done repeatedly until enough were obtained. Approximately 7-10 thrips were caged on each fruit within a plastic vial made of a 8 cm long by 5 cm diameter tube that had a very fine mesh organdy fabric glued to the bottom. The plastic cap was slit from the edge to the center so it could be slipped over the stem of the fruit. It was put in place, the thrips were knocked into the vial and it was fastened onto the cap then all gaps were closed with masking tape. The thrips naturally move up to the fruit. The cages were left in place for two days, then removed. The fruit were checked to be sure the thrips had stayed on, and then brought to the lab to cut off the flavedo using a razor blade. Only the flavedo from the stem 1/3 to 1/2 of the fruit was used. For controls an equal number of comparable sized fruit were caged without thrips, and the peel collected from them in the same manner. Tissues were frozen in liquid nitrogen, then stored at -80C until further processing. Fenton (Close lab) purified RNA by the phenol method described in J. Japanese Soc. Hort. Sci. 1996. 64 (4): 809-814, purified poly(A) mRNA using an Oligotex mRNA Kit (Qiagen), produced a primary cDNA library using a lambda ZAP XR cDNA Synthesis Kit (Stratagene), then mass-excised 0.77 million pfu from the primary library to produce a phagemid population. The library was made from equal portions of RNA from each of the two treatments. Phagemids were plated, and plasmid DNA purified by members of the 'Genomics & Biotechnology Laboratory' course (Duistermars, Bhakta, Resnik, Nguyen, Thanawiwat, Aromdee, Khan, Tsuji, Nguyen , Ikeda)taught by Close and Teaching Assistant Knoth. DNA sequences were determined bi-directionally using an ABI3730 at the UC Riverside Genomics Core Instrumentation Facility (Choi, Kingan). Chromatogram files were downloaded by FTP by Close, then processed by Wanamaker (Close lab) 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 assembled, then chimeras were removed following manual inspection of assemblies (Close, Roose, Federici, Wanamaker, Lyon, Ye, Jang, Collin, Kacar, Ikeda, Quinitio). Sequences that survived all removal steps were submitted to GenBank.

characteristic: description

text: 11 year old trees

text: E. coli TJC121

text: Citrus sinensis

text: EcoRI

text: XhoI

text: Flavedo

text: Parent Washington Navel Orange Thrip-Challenged Flavedo cDNA Library UCRCS07-153

text: Phagemid pBluescript SK(-)

text: Lambda Uni-ZAP XR, excised phagemid


text: USDA, ARS, National Center for Cool and Cold Water Aquaculture

text: live

text: Generic

text: WVU oocyte library.

text: Oncorhynchus mykiss

characteristic: organism

text: NCCCWA 10RT#1

characteristic: title

text: Plasmid

text: pCMV Sport6.0


text: Sun Yat-sen University School of Life Science

text: live

text: Generic

text: ESTs expressed differentially between Micronucleates and amicronucleates of Pseudourostyla cristata during starvation.

text: starvation

characteristic: dev-stage

text: Pseudourostyla cristata


text: live

text: Generic

text: WVU oocyte library RT-EGG.

text: Oncorhynchus mykiss

text: NCCCWA 10RT#2

text: Plasmid

text: pCMV Sport6.0


text: Department of Biology, University of Konstanz

text: live

text: Generic

text: stage 14

text: Pelodiscus sinensis

text: carapacial ridge

characteristic: tissue-type

text: Pelodiscus sinensis carapacial ridge stage 14


characteristic: INSDC center name

text: live

text: Generic

text: WVU oocyte library RT-EGG.

text: Oncorhynchus mykiss

text: NCCCWA 10RT#4

text: Plasmid

text: pCMV Sport6.0


text: Institut National de la Recherche Agronomique

text: live

text: Generic

text: Ugni blanc

text: Total RNA was isolated from 3, 6 and 15 days post anthesis for wild type and 'fleshless' mutated berries. cDNA was prepared from RNA (equal amount of the three developemntal stages for each genotype) using the smart PCR cDNA system (Clontech). cDNA library was performed with the PCR-select cDNA subtraction kit (BD Biosciences, ClonTech) using RNA from mutant as the tester and RNA from wild type as the driver.

text: Green stage

text: DH5 alpha

text: Fruit

characteristic: organ

text: Vitis vinifera

text: 'Fleshless' Ugni blanc SSH Library

text: pGem-T


characteristic: External Id

text: live

text: Generic

text: Ugni blanc

text: Total RNA was isolated from 3, 6 and 15 days post anthesis for wild type and 'fleshless' mutated berries. cDNA was prepared from RNA (equal amount of the three developemntal stages for each genotype) using the smart PCR cDNA system (Clontech). cDNA library was performed with the PCR-select cDNA subtraction kit (BD Biosciences, ClonTech) using RNA from wild type as the tester and RNA from mutant as the driver.

text: Green stage

text: DH5 alpha

text: Fruit

text: Vitis vinifera

text: Wild type Ugni blanc SSH Library

text: pGem-T


text: University of Georgia

text: live

text: Generic

text: Florunner

text: Arachis hypogaea

text: root

text: pPCR

text: Plasmid

text: pBluescript


text: University of Georgia

text: live

text: Generic

text: Florunner

text: Arachis hypogaea

text: stem

text: pPCS

text: Plasmid

text: pBluescript


text: University of Georgia

text: live

text: Generic

text: Florunner

text: Arachis hypogaea

text: root and stem

text: Kochert

text: Plasmid

text: pBluescript


text: live

text: Generic

text: WVU oocyte library RT-EGG.

text: Oncorhynchus mykiss

text: NCCCWA 10RT#3

text: Plasmid

text: pCMV Sport6.0


text: University of California

text: live

text: Generic

text: Parent Washington Navel

text: Trees were grown in the field at University of California, Riverside using standard horticultural practices. Young fruits were placed in a cage with thrips (Scirtothrips citri). Infestations were conducted by Watkins (Morse lab) and flavedo collected by Federici (Roose lab) from May-June 2003. The thrips were collected from naturally infested Rhus plants by sucking into a tube. A flexible hose was attached to a tube that extended into a covered vial, and another tube stuck out of the vial at a right angle. The bent tube was held above the thrips, sucking on the flexible tube created a vacuum, pulling the thrips into the vial. Thrips were knocked off the Rhus plant onto a manila folder, then only second instars were captured. This was done repeatedly until enough were obtained. Approximately 7-10 thrips were caged on each fruit within a plastic vial made of a 8 cm long by 5 cm diameter tube that had a very fine mesh organdy fabric glued to the bottom. The plastic cap was slit from the edge to the center so it could be slipped over the stem of the fruit. It was put in place, the thrips were knocked into the vial and it was fastened onto the cap then all gaps were closed with masking tape. The thrips naturally move up to the fruit. The cages were left in place for two days, then removed. The fruit were checked to be sure the thrips had stayed on, and then brought to the lab to cut off the flavedo using a razor blade. Only the flavedo from the stem 1/3 to 1/2 of the fruit was used. For controls an equal number of comparable sized fruit were caged without thrips, and the peel collected from them in the same manner. Tissues were frozen in liquid nitrogen, then stored at -80C until further processing. Fenton (Close lab) purified RNA by the phenol method described in J. Japanese Soc. Hort. Sci. 1996. 64 (4): 809-814, purified poly(A) mRNA using an Oligotex mRNA Kit (Qiagen), produced a primary cDNA library using a lambda ZAP XR cDNA Synthesis Kit (Stratagene), then mass-excised 0.77 million pfu from the primary library to produce a phagemid population. The library was made from equal portions of RNA from each of the two treatments. Phagemids were plated, plasmid DNA purified, cDNA clones archived, and DNA sequences determined bi-directionally using an ABI3730 at DNA Landmarks (Landry, Hubert, Laforest, Landry, Ligonde). Chromatogram files were downloaded by FTP by Close, then processed by Wanamaker (Close lab) 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 assembled, then chimeras were removed following manual inspection of assemblies (Close, Roose, Federici, Wanamaker, Lyon, Ye, Jang, Collin, Kacar, Ikeda, Quinitio). Sequences that survived all removal steps were submitted to GenBank.

text: 11 year old trees

text: E. coli TJC121

text: Citrus sinensis

text: EcoRI

text: XhoI

text: Flavedo

text: Parent Washington Navel Orange Thrip-Challenged Flavedo cDNA Library UCRCS07

text: Phagemid pBluescript SK(-)

text: Lambda Uni-ZAP XR, excised phagemid


text: University of California

text: live

text: Generic

text: Ruby

text: Two trees with open-pollinated flowers in a mixed planting in the UC Riverside Citrus Variety Collection were the source of tissue. Developing seeds were collected by Federici (Roose lab) from May-July 2003. This included nine stages, based on size of fruit: 10-20 mm, 20-30 mm, 30-35 mm, 35-40 mm, 40-45 mm, 45-50 mm, 50-55 mm, 55-60 mm, 60-65 mm. Tissues were stored in RNA Later (Ambion) until further processing. Fenton (Close lab) purified RNA by the phenol method described in J. Japanese Soc. Hort. Sci. 1996. 64 (4):809-814, purified poly(A) mRNA using an Oligotex mRNA Kit (Qiagen), produced a primary cDNA library using a lambda ZAP XR cDNA Synthesis Kit (Stratagene), then mass-excised 0.48 million pfu from the primary library to produce a phagemid population. The library was made from equal portions of RNA from seed at each of the nine fruit stages, but seeds were not of uniform size at any stage. Phagemids were plated, plasmid DNA purified, cDNA clones archived, and DNA sequences determined bi-directionally using an ABI3730 at DNA Landmarks (Landry, Hubert, Laforest, Landry, Ligonde). Chromatogram files were downloaded by FTP by Close, then processed by Wanamaker (Close lab) 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 assembled, then chimeras were removed following manual inspection of assemblies (Close, Roose, Federici, Wanamaker, Lyon, Ye, Jang, Collin, Kacar, Ikeda, Quinitio). Sequences that survived all removal steps were submitted to GenBank.

text: 20 year old trees

text: E. coli TJC121

text: Citrus sinensis

text: EcoRI

text: XhoI

text: Seed

text: Ruby Orange Developing Seed cDNA Library UCRCS09

text: Phagemid pBluescript SK(-)

text: Lambda Uni-ZAP XR, excised phagemid


text: Department of Animal Science, National Chung-Hsing University

text: live

text: Generic

text: Material with normal function was tested and heat stress experiments of animals were conducted by Divisions of Biotechnology and Animal Resources, Animal Technology Institute Taiwan (ATIT). First strand cDNA was primed with a NotI-oligo(dT) primer and 2nd strand cDNA was synthesized by using nick translational replacement of the mRNA. Double-strand cDNA was added with SalI adapter, then digested with NotI and cloned into the NotI and SalI sites of pSPORT1 vector.

text: adult, l year of age, heat stressed

text: DH10B (Life Technologies)

text: testis

text: Sus scrofa

text: NotI

text: SalI

text: male

characteristic: sex

text: Duroc

characteristic: strain

text: Porcine testis cDNA library II

text: Plasmid


text: University of Florida Food and Agricultural Sciences

text: live

text: Generic

text: Rubideaux

text: Cold-induced cDNA library generated from suppression subtractive hybridization of two day cold acclimated and non acclimated seedlings

text: One year old seedlings

text: Citrus trifoliata

text: RsaI

text: RsaI

text: Leaf

text: Cold-induced subtractive cDNA library

text: Plasmid

text: pT-ADV

characteristic: vector


text: Swiss Federal Research Station for Agroecology and Agriculture

characteristic: INSDC first public

text: live

text: Generic

text: Lolium multiflorum

text: young leaves

text: cDNA-AFLP LM-Xtg clones


text: USDA, ARS, National Center for Cool and Cold Water Aquaculture

characteristic: INSDC last update

text: live

text: Generic

text: This is an early neurogenesis SSH library created by Mathew L. Rise constructed by subtracting late neurogenesis (mixed stages: hindbrain swelling + heart tube with peristalsis) from early neurogenesis (mixed stages: neural groove + 1/2 epiboly). Fish were from a domesticated strain (Spring Valley Trout Farm, Langley, B.C.), courtesy of Bob Devlin, DFO. These are mostly internal (coding) sequences.

characteristic: lab-host

text: Oncorhynchus mykiss

text: NCCCWA 04RT

text: Plasmid

text: pCR 4-TOPO


text: University of California

text: live

characteristic: INSDC status

text: Generic

characteristic: NCBI submission model

text: Washington Navel

text: Parent Washington navel orange embryogenic callus was established from undeveloped ovules of ca. 10 mm diameter young fruits under open pollination on Murashige-Skoog medium at 25oC with 16 h light in a tissue culture room. Embryogenic callus, globular and heart stage embryoids were pooled in approximately equal portions in RNAlater (Ambion), then RNA was extracted using TRIZOL Reagent (Invitrogen). Poly(A) RNA was purified from 500 microgram of total RNA using Qiagen Oligotex. A primary cDNA library was produced using a lambda ZAP XR cDNA Synthesis Kit (Stratagene). These steps were performed by Xinrong Ye (Roose lab, UC Riverside). One million pfu from the primary library were mass excised to produce a phagemid population by Raymond Fenton (Close lab, UC Riverside). Phagemids were plated, plasmid DNA purified, cDNA clones archived, and DNA sequences determined bi-directionally using an ABI3730 at DNA Landmarks (Landry, Hubert, Laforest, Landry, Ligonde). Chromatogram files were downloaded by FTP to UC Riverside (by Close), then processed at UC Riverside (by Wanamaker, Close lab) 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 assembled, then chimeras were removed following manual inspection of assemblies (Close, Roose, Federici, Wanamaker, Lyon, Ye, Jang, Collin, Kacar, Ikeda, Quinitio). Sequences that survived all removal steps were submitted to GenBank.

text: Embryogenic and embryoid

text: E. coli TJC121

text: Citrus sinensis

text: EcoRI

text: XhoI

text: Callus

text: Parent Washington Navel Orange Callus cDNA Library UCRCS08-2

text: Phagemid pBluescript SK(-)

text: Lambda Uni-ZAP XR, excised phagemid


text: Washington University School of Medicine

text: live

text: Generic

characteristic: NCBI submission package

text: Arrayed normalized library of full-length cDNAs representing blastula stage transcriptome of the sea urchin Strongylocentrotus purpuratus, cloned into the vector pCMVSport6.1 (Invitrogen)

text: Strongylocentrotus purpuratus

text: Sea urchin EST Lib1

text: plasmid

text: pCMVSport6.1 (Invitrogen)


text: Institut National de la Recherche Agronomique

text: live

text: Generic

text: Gamay NJB

characteristic: cultivar

text: The SSH-cDNA fragments were generated using the SSH-adaptor sequences ctaatacgactcactatagggctcgagcggccgcccgggcaggt and ctaatacgactcactatagggcagcgtggtcgcggccgaggt. Total RNA was isolated from Gamay and Lacryma berries harvested 1 week after 'veraison'. cDNA was prepared from 2 ug of poly(A)+ RNA. cDNA library was performed with the PCR-select cDNA subtraction kit (BD Biosciences, ClonTech) using RNA from Gamay berries as the tester and RNA from Lacryma berries as the driver

characteristic: description

text: 1 week after 'veraison'

text: E. coli DH5 alpha competent cells

text: Fruit

text: Vitis vinifera

text: Gamay NJB cDNA Library

text: pGEM-T


text: live

text: Generic

text: Lacryma christii

text: The SSH-cDNA fragments were generated using the SSH-adaptor sequences ctaatacgactcactatagggctcgagcggccgcccgggcaggt and ctaatacgactcactatagggcagcgtggtcgcggccgaggt. Total RNA was isolated from Gamay and Lacryma berries harvested 1 week after 'veraison'. cDNA was prepared from 2 ug of poly(A)+ RNA. cDNA library was performed with the PCR-select cDNA subtraction kit (BD Biosciences, ClonTech) using RNA from Lacryma berries as the tester and RNA from Gamay berries as the driver

text: 1 week after 'veraison'

text: E. coli DH5 alpha competent cells

text: Fruit

text: Vitis vinifera

characteristic: organism

text: Lacryma berries cDNA Library

characteristic: title

text: pGEM-T


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

text: live

text: Generic

text: Entamoeba histolytica

text: E. histolytica Normalized cDNA library


text: National Centre for Plant Genome Research

text: live

text: Generic

text: Prabal

text: One year old plant

characteristic: dev-stage

text: Catharanthus roseus

text: Leaf specific cDNA library of one year old Catharanthus roseus var. Prabal


text: Hanyang University

characteristic: INSDC center name

text: live

text: Generic

text: Light stressed

characteristic: cell type

text: High Light stressed Dunaliella, Uni-ZAP XR Vector System(Stratagene),

text: vegetative, mid-log phase growing stage

text: SOLR (E. coli strain)

text: Dunaliella salina

text: EcoRI

text: XhoI

text: UTEX 1644

text: High Light stressed Dunaliella salina

text: phagemid

text: pBluescript SK-


characteristic: External Id

text: Institution of Biotechnology

text: live

text: Generic

text: epithelial and mesenchymal cells

text: Amplified nanoquantity cDNA library prepared by ResGen / Invitrogen.

text: Embryonic day 10

text: pancreas

characteristic: organ

text: Mus musculus

text: Female and male, pooled

text: C57BL/6xCBA F1 hybrids

text: pancreas

characteristic: tissue-type

text: Embryonic day 10 Mouse Pancreas Amplified cDNA library

text: pENTR 222


text: Department of Biotechnology

text: live

text: Generic

text: An oligo(dT) primed library was constructed in pCMVsport6 from RNA isolated from lateral ventricle wall tissue of adult male and female mice.

text: Adult

text: NeuroNova AB

text: Brain

text: Mus musculus

text: NotI

text: EcoRV

text: Male and Female

characteristic: sex

characteristic: strain

text: Lateral Ventricle Wall

text: Mus Musculus Lateral Ventricle Wall C57BL/6 adult

text: Plasmid

characteristic: vector


text: Laval University

characteristic: INSDC first public

characteristic: INSDC last update

text: live

text: Generic

text: Oocytes

text: T/A cloning with pCR4-TOPO vector from Invitrogen; Library made by Suppressive Subtractive Hybridization from bovine oocytes.

text: Ovary

text: Bos taurus

text: female

text: Bovine oocyte cDNA subtracted library

text: pCR4-TOPO vector


text: USDA, ARS, National Center for Cool and Cold Water Aquaculture

text: live

characteristic: INSDC status

text: Generic

characteristic: NCBI submission model

text: This is a late neurogenesis SSH library created by Mathew L. Rise by subtracting early neurogenesis (mixed stages: neural groove + 1/2 epiboly) from late neurogenesis (mixed stages: hindbrain swelling + heart tube with peristalsis). Fish were from a domesticated strain (Spring Valley Trout Farm, Langley, B.C.), courtesy of Bob Devlin, DFO. These are mostly internal (coding) sequences.

characteristic: lab-host

text: Oncorhynchus mykiss

text: NCCCWA 05RT

text: Plasmid

text: pCR 4-TOPO


text: Departement de Biochimie, Universite de Montreal

text: live

text: Generic

characteristic: NCBI submission package

text: Helicosporidium sp. ex Simulium jonesi

text: Helicosporidium sp. Non-normalized


text: University of Waterloo

text: live

text: Generic

text: Yukon

characteristic: cultivar

text: Library was prepared following the protocol for Invitrogen SuperScript Plasmid System with GAteway Technology for cDNA synthesis and cloning

characteristic: description

text: 4-week old plants

text: Eutrema salsugineum

characteristic: organism

text: Leave

text: Cold Library

text: plasmid


text: New Mexico State University

characteristic: INSDC center name

text: live

text: Generic

text: drought-stressed

characteristic: dev-stage

text: root

text: Phaseolus vulgaris

text: Xho 1

text: negro jamapa bean root cDNA

characteristic: title

text: Phage

text: Zap


characteristic: External Id

text: New Mexico State University

text: live

text: Generic

text: drought-stressed

text: Root

characteristic: organ

text: Phaseolus acutifolius

text: Xho 1

text: tepary bean root cDNA

text: Phage

text: Zap


text: Faculty of Veterinary Medicine, Universite de Montreal

text: live

text: Generic

text: PCR fragment; cDNA library made from intact skin that was subtracted from wound margin biopsies obtained 7 days post wounding.

text: proliferative phase of wound healing

text: skin

text: Equus caballus

text: female

characteristic: sex

text: Equus caballus subtracted cDNA of intact skin from wound margin biopsies (WM-IS).


text: live

text: Generic

text: This is a normalized (Cot = 5) O. mykiss whole juvenile library created by Matthew L. Rise from approximately 4 month old O. mykiss (Tzenzaicut Lake strain) obtained from Vancouver Island Trout Hatchery (Duncan, B.C.).

text: Oncorhynchus mykiss

text: pooled

characteristic: tissue-type

text: NCCCWA 03RT

text: Plasmid

text: pBluescript SK+


text: Chang Gung University

text: live

text: Generic

characteristic: cell-line

text: Unsynchronized mid-log phase culture

text: XL1 Blue-MRF'

text: Tritrichomonas foetus

text: EcoRI

text: XhoI

text: Tf30924 cDNA Library

text: Lambda ZAP-Express (Stratagene)

characteristic: vector


text: Lawrence Berkeley National Lab

characteristic: INSDC first public

characteristic: INSDC last update

text: live

text: Generic

text: Sized fractionated cDNAs were directly ligated into pOTB7. Plasmid cDNA library.

text: 0 to 24 hours mixed stage embryonic

text: embryo

text: Drosophila pseudoobscura

text: EcoRI

text: male and female

text: DP Drosophila pseudoobscura Embryo


text: Lawrence Berkeley National Lab

text: live

characteristic: INSDC status

text: Generic

characteristic: NCBI submission model

text: 0 to 24 hours mixed stage embryonic

text: embryo

text: Drosophila virilis

text: EcoRI

text: male and female

text: DV Drosophila virilis Embryo


text: c/o UniMi DiProVe, Parco Tecnologico Padano

text: live

text: Generic

characteristic: NCBI submission package

text: endocarp hardening

text: Prunus persica

text: fruit mesocarp


text: live

text: Generic

text: post-fertilization

text: Prunus persica

text: fruit mesocarp


text: Iowa State University

text: live

text: Generic

text: Inbred B73

characteristic: cultivar

text: This library was constructed by Xiaolan Zhang. Vegetative Shoot Apical Meristem (SAM) and leaf primordia staged P1-P4 from 14-17 day-after germination seedlings were quickly dissected into dry ice under a light microscope. Total RNA was isolated using Trizol and mRNA was purified with Dynal Oligo-DT25. ds-cDNA molecules were generated as follows. First-strand cDNA was prepared from oligo-dT selected mRNA by priming with an XhoI oligo-dT primer (5'-GAGAGAGAGAGAGAGAGAGAACTAGTCTCGAGTTTTTTTTTTTTTTTTTT). The resulting DNA:RNA hybrid was treated with RNase H and used as a template for DNA PolI-catalyzed second strand synthesis. After the addition of EcoRI adaptors, the ds-cDNAs were digested with XhoI and size-selected to be >600 bp. The resulting molecules were directionally cloned into the EcoRI and XhoI sites of the Uni-Zap XR vector. The lambda library was packaged with Gigapack III Gold packaging extract and was mass excised by XL1-Blue cells and ExAssist helper phage. Excised phagemids were titered in SOLR cells and plated onto LB-ampicillin agar plates. Base calling was conducted using Phred. Trimming was performed using Lucy with the following criteria: (-minimum 200 -error 0.01 0.01 -bracket 10 0.01). A low complexity filter was applied and additional trimming was conducted to remove E. coli, vector, and organelle contamination. After processing ~30% of the sequences contained a minimum of 10 Ts at the beginning of the sequence. For reasons that are not understood many of the clones in this library lack an XhoI site at their 3' ends.

characteristic: lab-host

text: Shoot apex

text: Zea mays

text: EcoRI

text: XhoI

text: Uni-Zap XR


text: Nanjing Forestry University

text: live

text: Generic

text: Populus deltoides

characteristic: organism

text: EcoRI

text: XhoI

text: leaf inoculated with Marssonia pathogen of Populus deltoides

text: Phage

text: pBK-CMV


text: Bielefeld University, Institute for Genome Research

characteristic: INSDC center name

text: live

text: Generic

text: Express 617

text: cDNA library from Brassica napus, strain Express 617; RNA was from young flowers and flower buds of two greenhouse plants; library was made at the Max-Planck-Institute for Plant Breeding Research, Cologne, Germany; cloning sites SalI-NotI, primer sites and orientation: T7-SalI-CCACGCGTCCG-5str-cDNA-polyA-CC-NotI-SP6; made by Marc Jakoby 09/2000; PI: Bernd Weisshaar. Sequence submission managed by RZPD/GABI-Primary database: http://gabi.rzpd.de

characteristic: description

text: young flowers and flower buds

text: Brassica napus

text: SalI

text: NotI

text: flowers and flower buds

text: ADIS-MPIZ 021


text: Texas Agricultural Experiment Station

text: live

text: Generic

text: 3-week old seedlings

characteristic: dev-stage

text: E. coli JM109

text: Aegilops tauschii

text: leaves and stems

text: cDNA-AFLP clones from Aegilops tauschii

characteristic: title

text: pGEM-T Easy


characteristic: External Id

text: Texas Agricultural Experiment Station

text: live

text: Generic

text: 3-week old seedlings

text: Triticum aestivum

text: leaves and stems

text: cDNA-AFLP clones from wheat

text: pGEM


text: live

text: Generic

text: This library was created by A.-S. Goupil and Y. Guiguen who subtracted the NCCCWA 1RT library from the INRA multi-tissue library.

text: Oncorhynchus mykiss

text: pooled

text: NCCCWA 02RT

text: Plasmid

text: pCMV Sport6.0


text: Northeast Forestry University

text: live

text: Generic

text: Limonium bicolor

text: cDNA library of Limonium bicolor (Bunge) Kuntze


text: Royal Veterinary and Agricultural University

text: live

text: Generic

text: FMV-1 (IE 4673)

text: mRNA was isolated from seeds after removal of seed coat. Developmental stages range from 1st appearance of starchy endosperm to onset of seed desiccation. Clones for sequencing were produced from unamplified library. Average insert size is 1.1kbp.

text: Developing

text: Eleusine coracana

text: SalI (5' end of cDNA)

text: NotI (3' end of cDNA)

text: Seed (Endosperm, Embryo, Testa)

text: Finger millet developing seed cDNA

text: Plasmid


text: National Research Council of Canada

text: live

text: Generic

text: Brassica rapa 39RDBRT. Sequences have been trimmed to remove vector and low quality regions using LUCY sequence cleanup software (www.tigr.org).

text: Brassica rapa

text: Sfi IA

text: Sfi IB

text: Brassica rapa 39RDBRT

text: plasmid


text: National Research Council of Canada

text: live

text: Generic

text: Brassica rapa 38RDBRM. Sequences have been trimmed to remove vector and low quality regions using LUCY sequence cleanup software (www.tigr.org).

text: Brassica rapa

text: Sfi IA

text: Sfi IB

text: Brassica rapa 38RDBRM

text: plasmid


text: National Research Council of Canada

text: live

text: Generic

text: Brassica napus 21ACACMS. Sequences have been trimmed to remove vector and low quality regions using LUCY sequence cleanup software (www.tigr.org).

text: Brassica napus

text: Sfi IA

text: Sfi IB

text: Brassica napus 21ACACMS

text: plasmid


text: National Research Council of Canada

text: live

text: Generic

text: Germinating seeds 4h. Sequences have been trimmed to remove vector and low quality regions using LUCY sequence cleanup software (www.tigr.org).

text: Brassica napus

text: Germinating seeds 4h

text: plasmid


text: National Research Council of Canada

text: live

text: Generic

text: Brassica napus 16ACDHDS. Sequences have been trimmed to remove vector and low quality regions using LUCY sequence cleanup software (www.tigr.org).

text: Brassica napus

text: Sfi IA

text: Sfi IB

text: Brassica napus 16ACDHDS

text: plasmid


text: National Research Council of Canada

text: live

text: Generic

text: Brassica napus 20ACACDS. Sequences have been trimmed to remove vector and low quality regions using LUCY sequence cleanup software (www.tigr.org).

text: Brassica napus

text: Sfi IA

text: Sfi IB

text: Brassica napus 20ACACDS

text: plasmid


text: National Fisheries Research & Development Institute

text: live

text: Generic

text: Adult

text: E.coli SOLR

text: Paralichthys olivaceus

text: EcoR I

text: Xho I

text: Paralichthys olivaceus cDNA library

text: Phagemid vector

text: pBluescript SK(-)


text: Instituto de Biologia Molecular y Celular de Plantas (Universidad Politecnica de Valencia - Consejo Superior de Investigaciones Cientificas)

text: live

text: Generic

text: cDNA library made from leaves of plants infected with Citrus Tristeza Virus (CTV)

text: adult trees

text: Escherichia coli

text: leaves

characteristic: organ

text: Citrus macrophylla

text: hermaphrodite

characteristic: sex

text: Plasmid


text: live

text: Generic

text: Cleopatra

text: Subtracted cDNA library made from a mixture of equal amounts of poly-A+ RNA from roots of plants subjected to 5, 10, and 24 hours of drought stress, subtracted with poly-A+ RNA from roots of unstressed plants

text: two-year old plants

text: Escherichia coli

text: roots

text: Citrus reshni

text: hermaphrodite

text: Plasmid


text: live

text: Generic

text: Cidro Etrog selection 861-S1

text: cDNA library made from leaves of plants showing symptoms of viroid infection 3 months after inoculation with an Citrus exocortis viroid (CEVd) isolate by shoot incision

text: adult trees

text: Escherichia coli

text: leaves

text: Citrus medica

text: hermaphrodite

text: Plasmid

text: Lambda-ZAP II


text: live

text: Generic

text: Carrizo Citrange

text: cDNA library made from a mixture of equal amounts of poly-A+ RNA from roots of plantas showing growing chlorosis ferric symptoms at 4 different flushing along one year of iron-deficiency treatment (by transplanting plants growing in standard greenhouse conditions to pots containing calcareous soil (70% CaCO3), where iron, in Fe+++ form, is not available for the plant)

text: adult trees

text: Escherichia coli

text: Roots

text: Citrus sinensis x Citrus trifoliata

text: hermaphrodite

text: Plasmid

text: Lamda-ZAP


text: live

text: Generic

text: Fortune

text: cDNA library made from a mixture of equal amounts of poly-A+ RNA from flavedo of stored fruits heat-treated at 37oC for 4, 24, and 72 hours, and of fruits heat-treated at 37oC for 72 hours and then stored at 1oC for 1, 3, and 10 days

text: adult trees

text: Escherichia coli

text: fruits

text: Citrus clementina x Citrus tangerina

text: hermaphrodite

text: flavedo

characteristic: tissue-type

text: Plasmid

text: Uni-ZAP XR


text: live

text: Generic

text: Fortune

text: Subtracted cDNA library made from poly-A+ RNA from flavedo of fruits stored at 2oC for 14 days, subtracted with poly-A+ RNA from fruits stored at 12oC for 14 days

text: adult trees

text: Escherichia coli

text: fruits

text: Citrus clementina x Citrus tangerina

text: hermaphrodite

text: flavedo

text: Plasmid

characteristic: vector


characteristic: INSDC first public

text: live

text: Generic

text: cDNA library made from a mixture of equal amounts of poly-A+ RNA from roots of plant inoculated with Phytophthora citrophtora zoospores by inmersion in a suspension with 2000 or 15000 zoospores/mL, extracted at 0, 2, 4, 8, 13, and 18 days after inoculation

text: 2-3 month old seedlings, 20-30 cm. high

text: Escherichia coli

text: roots

text: Citrus sinensis

text: hermaphrodite

text: Plasmid

text: Uni-ZAP XR


characteristic: INSDC last update

text: live

text: Generic

text: Carrizo Citrange

text: cDNA library made from poly-A+ RNA from 0.5 cm. root tips of seedlings grown in Paclobutrazol (giberellin synthesis inhibitor) or auxin (indol-butyric acid) for 1-4 days

text: seedlings

text: Escherichia coli

text: roots

text: Citrus sinensis x Citrus trifoliata

text: hermaphrodite

text: Plasmid

text: Lambda-ZAP II


text: Lawrence Berkeley National Lab

text: live

characteristic: INSDC status

text: Generic

text: RT-PCR products were amplified with pairs of gene-specific primers from template RNA representing pooled Drosophila stages. Products were directly sequenced from both ends with the same oligos used to prime PCR.

text: 0 to 24 hours mixed stage embryonic, late third instar larvae, mixed stage pupae, mixed age adults

text: Drosophila melanogaster

text: male and female

text: AM Drosophila melanogaster Embryo Larva Pupa Adult


text: USDA, ARS, NADC

text: live

text: Generic

characteristic: NCBI submission model

text: The Lymph node SSH library is derived from the suppression subtractive hybridization between Salmonella enterica subsp. enterica serotype Choleraesuis infected and non infected porcine lymph node.

text: DH5a (Life Technologies)

text: Sus scrofa

text: crossbreed

characteristic: strain

text: Lymph Node

text: Lymph-node-SSH

text: pBAD (Invitrogen)


text: VBI

text: live

text: Generic

characteristic: NCBI submission package

text: (Adopted from the Invitrogen product manual) Custom cDNA Library pCMV-SPORT6.1 is prepared as follows: mRNA is isolated using two steps. First, total RNA is isolated from tissues or cells using the TRIzol Reagent. Second, mRNA is isolated from total RNA using oligo (dT) in a filter syringe. First-strand cDNA is synthesized using SuperScript III Reverse Transcriptase Second-strand cDNA is synthesized using E. coli RNase H, E. coli DNA polymerase I, and E. coli DNA ligase cDNA is blunt-ended using T4 DNA Polymerase and digested with Not I cDNA is size-selected using column chromatography or agarose gel electrophoresis Size-selected cDNA is directionally cloned into the Not I-EcoR V region of the vector (EcoR V site is destroyed during cloning) Ligation mixture is transformed into competent DH10B T1-Phage Resistant E. coli and the number of primary recombinants is determined

text: seedlings

text: Fragaria vesca

text: NotI

text: EcoRV

text: Fragaria vesca heat stressed seedlings cDNA library in pCMV-SPORT 6.1

text: Plasmids


text: Peking University

text: live

text: Generic

text: preimplantation embryos

text: Mus musculus

text: Mus musculus preimplantation embryos


text: Shanghai Institute of Animal Parasitology, Chinese Academy of Agricultural Sciences

text: live

text: Generic

text: Cercaria subtracted cDNA library using cDNA from cercaria as testers and from adult worm as drivers. Cercaria cDNAs were synthesized from 1ug total RNA using SMART PCR cDNA Synthesis Kit according to the manufacturer's protocol ( clontech, palo Alto, CA ), because cercaria is very limited. Subtractive PCRs were performed using the PCR-select cDNA subtractive kit according to the manufacture's protocol (clontech, palo Alto, CA) with the following modification. Xho I site and EcoR I site were respectively added to the 5' end of nested PCR primer 1 and nested PCR primer 2R. Sequence as follows: nested PCR primer 1 5' AGTTCTCGAGCGGCCGCCC GGGCAGGT 3', nested PCR primer 2R 5' TCGGGAATTCTGGTCGGTCGCGG CCGAGGT 3'. Thus PCR products can be directionally subcloned into the pCMV-SCRIPT ( XR ) vector ( stratagene ) and transformed into XL1-Blue electrophoration-competent cells ( stratagene ).

text: Larva

text: Rabbit

characteristic: lab-host

text: Schistosoma japonicum

text: EcoRI

text: XhoI

text: Mix

text: Chinese mainland

text: Whole body

text: Cercaria subtracted cDNA library

text: Plasmid

text: pCMV-SCRIPT