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Library Info Page


Library ID

Library Name:NIA Mouse E10.5 whole embryo cDNA library (Long)
Organism:Mus musculus
Strain:C57BL/6J
UniGene Library ID:15706
Tissue Description:Embryo, mouse E10.5 whole embryo including extraembryonic tissues, pool of 8
Library Keywords:normal, bulk, adult, long-transcript enriched, EST, mixed tissue, MGC, embryonic tissue, PCR directed strategy, cDNA template, mixed developmental stages, extraembryonic tissue, plasmid vector, oligo-dT primed, C57BL/6J, organogenesis

Clones and Sequences

#Clones Generated to Date:9600
#Sequences Generated to Date:7091

Library Preparation Details

Description:Mouse cDNA project by the Laboratory of Genetics, National Institute on Aging (NIA), Intramural Research Program, NIH (http://lgsun.grc.nia.nih.gov/cDNA). This is a long-transcript enriched cDNA library (Ref. Genome Res. 11: 1553-1558 (2001). [PMID: 11544199]). Total RNAs were extracted from a pool of 8 embryos at 10.5-days postcoitum. Double-stranded cDNAs were synthesized with an Oligo(dT) primer [Invitrogen: 5'-pGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'] from 2ug of total RNA, treated with T4 DNA polymerase, and purified by ethanol-precipitation. The cDNAs were ligated to Lone-linker LL-Sal4, purified by phenol/chloroform, and separated from free linkers by Centricon 100. Then, the cDNAs were amplified by long-range high fidelity PCR using Ex Taq polymerase (Takara) with a primer Sal4-S. The products were purified by phenol/chloroform and Centricon 100. The cDNAs were digested with SalI and NotI enzymes and cloned into SalI/NotI site of pCMV-SPORT6 plasmid vector. The DH10B E. coli host was transformed with the ligation mixture by the standard chemical method. The average insert size is about 3.4Kb. The library was constructed by Yulan Piao.
R. Site1:SalI
R. Site2:NotI
Lab Host:DH10B
Vector:pCMV-SPORT6 (Invitrogen)
Vector Type:plasmid (ampicillin resistant)
Tissue Supplier:Minoru Ko, NIA
Library Producer:Yulan Piao and Minoru Ko (National Institute on Aging, NIH: http://lgsun.grc.nia.nih.gov/cDNA/)