Name: NIA Mouse ES Cell (LIF-) cDNA Library (Long) Library ID: 1991 Organism: Mus musculus Strain: 129/Sv x 129S1/Sv-p<+> Tyr-c<+> Age: 0 Stage: embryo Organ: embryonic stem cell Tissue: Embryonic Stem Cell (LIF-) 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 absence of LIF for 4 or 18 hours. Equimolar mixtures of these RNA samples were used for library construction. Double- stranded cDNAs were synthesized with an oligo-dT primer (5'- pGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'), 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].
