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Cancer Genome Characterization Initiative

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Library Info Page


Library ID

Library Name:Stanley Frontal SB pool 1
Organism:Homo sapiens
UniGene Library ID:1478
Tissue Description:
Library Keywords:frontal cortex, bulk, adult, subtracted, EST, male, size fractionated, plasmid vector, oligo-dT primed

Clones and Sequences

#Clones Generated to Date:
#Sequences Generated to Date:205

Library Preparation Details

Description:Total RNA (purified with Trizol and DNAsed before use) was reverse transcribed using a modified oligo-dT primer containing RsaI and HindIII sites. Double- stranded cDNA was digested with RsaI, resulting in blunt ended cDNA of an average 0.1-2 kb in length. Digested cDNA was split into two sets, one used as is as the driver, the other set was split in half again and each half linked to a different adaptor (5'-TCGAGCGGCCGCCCGGGCAGGT-3' or 5'- AGGGCGTGGTGCGGAGGGCGGT-3'), to be used as tester. Subtraction was performed using the Clontech PCR Select cDNA subtraction kit. Pool of two male schizophrenics, ages 30 and 32 (S-30, S-121) subtracted by pool of two male bipolars, age 34 and 48 (S-75, S-83). Tissues were obtained from the Stanley Neuropathology Consortium (www.stanleylab.org). Library constructed and subtracted by Dr. Nancy Johnston [(410) 614-3918, nlj@welchlink.welch.jhu.edu].
R. Site1:EcoRI
R. Site2:
Lab Host:DH10B (phage-resistant)
Vector:pCR2.1-TOPO (Invitrogen)
Vector Type:plasmid (ampicillin resistant)
Tissue Supplier:
Library Producer: