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Cancer Genome Characterization Initiative

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Library Info Page

Library ID

Library Name:NIA Mouse Embryonic Stem (ES) cell (Lif-, 48 h, low density) cDNA library (Long)
Organism:Mus musculus
UniGene Library ID:15704
Tissue Description:Embryonic Stem (ES) cell, mouse, Lif-, 48 h, low density (129.3 ES cell line)
Library Keywords:normal, cell line, long-transcript enriched, EST, embryonic stem cell, MGC, embryonic tissue, male, unknown embryonic stage, PCR directed strategy, cDNA template, plasmid vector, oligo-dT primed

Clones and Sequences

#Clones Generated to Date:9600
#Sequences Generated to Date:7210

Library Preparation Details

Description:Mouse cDNA project by the Laboratory of Genetics, National Institute on Aging (NIA), Intramural Research Program, NIH ( This is a long-transcript enriched cDNA library (Ref. Genome Res. 11: 1553-1558 (2001). [PMID: 11544199]). ES cells were plated at density 3x103/cm2, on gelatin-coated plates and cultured for 48 hrs at 37 OC, 5% CO2. Culture medium: DMEM supplemented with 15% FBS, 2 mM L-glutamine, 0.1 mM NEAA, 1mM Sodium pyruvate, 0.1 mM beta-mercaptoethanol, 100 U/ml penicillin, and 100 ug/ml streptomycin. Double-stranded cDNAs were synthesized with an Oligo(dT) primer [Invitrogen: 5'-pGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'] from 2ug of total RNA, treated with T4 DNA polymerase, and purified by ethanol-precipitation. The cDNAs were ligated to Lone-linker LL-Sal4, purified by phenol/chloroform, and separated from free linkers by Centricon 100. Then, the cDNAs were amplified by long-range high fidelity PCR using Ex Taq polymerase (Takara) with a primer Sal4-S. The products were purified by phenol/chloroform and Centricon 100. The cDNAs were digested with SalI and NotI enzymes and cloned into SalI/NotI site of pCMV-SPORT6 plasmid vector. The DH10B E. coli host was transformed with the ligation mixture by the standard chemical method. The average insert size is about 2.8 kb. The library was constructed by Yulan Piao.
R. Site1:SalI
R. Site2:NotI
Lab Host:DH10B
Vector:pCMV-SPORT6 (Invitrogen)
Vector Type:plasmid (ampicillin resistant)
Tissue Supplier:Minoru Ko, NIA
Library Producer:Yulan Piao and Minoru Ko (National Institute on Aging, NIH: