Name: NIA Mouse Hematopoietic Stem Cell (Lin-/c-Kit+/Sca-1-) cDNA Library (Long) Library ID: 2012 Organism: Mus musculus Strain: C57BL/6NCr Age: 0 Stage: juvenile Organ: stem cell Tissue: hematopoietic stem cell (Lin-/c-Kit+/Sca-1-) 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 Drs. Dennis Taub, Dan Longo (National Institute on Aging, USA) and Jonathan Keller (National Cancer Institute, USA). Double-stranded cDNAs were synthesized with an oligo-dT primer (5'- pGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3')from 2.4 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 2.2 kb. The library was constructed by Yulan Piao and Minoru Ko (NIH, NIA). Reference: Genome Res (2001) 11:1553-1558 [PMID: 11544199].
