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cDNA library: NIA Mouse Undifferentiated Embryonic Stem (ES) Cell cDNA Library (Long 1)

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embryonic stem cell

   1. NIA Mouse Undifferentiated Embryonic Stem (ES) Cell cDNA Library (Long 1)

NIA Mouse Undifferentiated Embryonic Stem (ES) Cell cDNA Library (Long 1)

Name: NIA Mouse Undifferentiated Embryonic Stem (ES) Cell cDNA Library (Long 1)
Library ID: 2102
Organism: Mus musculus
Strain: 129/Sv x 129S1/Sv-p<+> Tyr-c<+>
Age: 0
Organ: embryonic stem cell
Tissue: undifferentiated embryonic stem cells
Host: DH10B
Vector: pCMV-SPORT6
Vector type: phagemid
Insert digest: 5' SalI/NotI 3'
Stop Codon Status: without
Description: Mouse cDNA project by the Laboratory of Genetics, National Institute on Aging (NIA), Intramural Research Program, NIH (http://lgsun.grc.nia.nih.gov/cDNA). This is a long-transcript enriched cDNA library (Ref. Genome Res. 11: 1553-1558 (2001). [PMID: 11544199]). Total RNAs were obtained from Dr. Kenneth R. Boheler (National Institute on Aging, USA). ES cells were cultured without feeder cells in the presence of LIF and BRL-conditioned media. Double-stranded cDNAs were synthesized with an Oligo(dT) primer [Invitrogen: 5'-pGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'] from 14.2 5g 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 2.4 kb. The library was constructed by Yulan Piao.

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