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Cancer Genome Characterization Initiative

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Library Info Page


Library ID

Library Name:NIH_MGC_151
Organism:Homo sapiens
UniGene Library ID:15410
Tissue Description:Breast, human breast cancer cells growing in a xenograft mouse model
Library Keywords:mammary gland, non-normalized, flow-sorted, EST, unknown developmental stage, MGC, female, xenograft, size fractionated, phagemid, oligo-dT primed, breast (mammary gland) cancer

Clones and Sequences

#Clones Generated to Date:4992
#Sequences Generated to Date:117

Library Preparation Details

Description:Cells were isolated by flow cytometry and RNA isolated in Trizol reagent. cDNA was obtained by reverse transcription using oligo-dT primer 5'-GACCACGCGTATCGATGTCGACTTTTTTTTTTTTTTTTV-3'and treated with ExoI. PCR was performed using a complementary 3' primer containing a BstXI site and a cap-switch 5' primer. The resulting cDNA was digested with BstXI and ligated to a BstXI adaptor 5'-AAAAAAAAAAAAAAAAGTCGACATCGATACGCGTGGTCTCGTAGCTGACTTTCCAGCACA-3' then digested with AscI. Size-selection was performed >0.5 kb to result in an average insert size of 0.8 kb. Library constructed and contributed by Mike Clarke and Andrew Hass (University of Michigan). Note: this is an MGC library.
R. Site1:AscI
R. Site2:BstXI
Lab Host:DH10B
Vector:pOTB7-3
Vector Type:phagemid
Tissue Supplier:
Library Producer:Mike Clarke , Angie Parks