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cDNA library: NIA Mouse Hematopoietic Stem Cell (Lin-/c-Kit-/Sca-1+) cDNA Library (Long)

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   1. NIA Mouse Hematopoietic Stem Cell (Lin-/c-Kit-/Sca-1+) cDNA Library (Long)

NIA Mouse Hematopoietic Stem Cell (Lin-/c-Kit-/Sca-1+) cDNA Library (Long)

Name: NIA Mouse Hematopoietic Stem Cell (Lin-/c-Kit-/Sca-1+) cDNA Library (Long)
Library ID: 2014
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 1.1 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].

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