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Library Info Page


Library ID

Library Name:NIA Mouse Trophoblast Stem Cell cDNA Library (Short)
Organism:Mus musculus
Strain:B5/EGFP transgenic ICR mice
UniGene Library ID:10025
Tissue Description:
Library Keywords:normal, cell line, pregnant, adult, short-transcript enriched, EST, female, PCR directed strategy, cDNA template, trophoblast stem cell, plasmid vector, oligo-dT primed, trophoblast

Clones and Sequences

#Clones Generated to Date:
#Sequences Generated to Date:1538

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 short-transcript enriched cDNA library (Ref. Genome Res. 11: 1553-1558 (2001). [PMID: 11544199]). Total RNAs were obtained from Dr. Janet Rossant and Tilo Kunath (Samuel Lunenfeld Research Institute, Canada). Double-stranded cDNAs were synthesized with an Oligo(dT) primer [Invitrogen: 5'-pGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'] from 4 ug 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-L. 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 pSPORT1 plasmid vector. The DH10B E. coli host was transformed with the ligation mixture by the standard chemical method. The average insert size is about 1.3 kb. The library was constructed by Yulan Piao (NIA).
R. Site1:SalI
R. Site2:NotI
Lab Host:DH10B
Vector:pSPORT1 (Invitrogen)
Vector Type:plasmid (ampicillin resistant)
Tissue Supplier:
Library Producer: