BioSample

Mouse cDNA project by the Laboratory of Genetics, National Institute on Aging (NIA), Intramural Research Program, NIH (http://lgsun.grc.nia.nih.gov/cDNA). Double-stranded cDNAs were synthesized with an Oligo(dT) primer [Invitrogen: 5'- pGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'] from 48 microgram of total RNA, treated with T4 DNA polymerase, and purified by ethanol-precipitation. The cDNAs were ligated to Lone-linker LL-Sal3 (Ref. Development 127:1737-1749 (2000) [PMID:10725249]), purified by phenol/chloroform, and separated from free linkers by Centricon 100. Then, the cDNAs were digested with SalI and NotI enzymes, and cloned into SalI/NotI site of pSPORT1 plasmid vector. The DH10B E. Coli host was trasnformed with ligation mixture by the standard chemical method. The average insert size is about 1.9 kb. The library was constructed by Yulan Piao (NIA).