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

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Information for 1 library

embryonic stem cell

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

NIA Mouse Undifferentiated ES Cell cDNA Library (Long)

Name: NIA Mouse Undifferentiated ES Cell cDNA Library (Long)
Library ID: 1992
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: pSPORT1
Vector type: phagemid
Insert digest: 5' SalI/NotI 3'
Stop Codon Status: without
Description: This is a long-transcript enriched cDNA library. Total RNA was obtained from Dr. Kenneth R. Boheler (National Insitute 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 (5'- pGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3') from 14.2 ug of total RNA, treated with T4 DNA polymerase, and purified by ethanol precipitation. The cDNAs were ligated to Lone-linker LL-Sal4 (5'-AAGAGGTAATCC-3'), purified by phenol/chloroform, and separated from free linker by Centricon 100. The cDNAs were amplified by long-range high fidelity PCR using Ex Taq polymerase (Takara) with primer Sal4-S. The cDNAs were digested with SalI and Not1I enzymes and cloned into SalI/NotI sites of pSPORT1 plasmid vector. The average insert size is about 2.4 kb. The library was constructed by Yulan Piao and Minoru Ko (NIH, NIA). Reference: Genome Res (2001) 11:1553-1558 [PMID: 11544199].

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