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Cancer Genome Characterization Initiative

Visit the database of genomic characterization data for multiple tumor types.

Library Info Page

Library ID

Library Name:Myeloma (MYE) cDNA library
Organism:Homo sapiens
UniGene Library ID:7038
Tissue Description:
Library Keywords:bone marrow, non-normalized, flow-sorted, multiple myeloma, EST, unknown developmental stage, male, size fractionated, phage vector, oligo-dT primed

Clones and Sequences

#Clones Generated to Date:
#Sequences Generated to Date:3676

Library Preparation Details

Description:Myeloma cells from multiple myeloma patients' bone marrow were purified by magnetic cell sorting. mRNA were purified and an oligo d(T)18 primer containing XhoI restriction site was used to prime first strand synthesis using M-MLV reverse transcriptase. To protect the cDNAs from XhoI digestion in subsequent cloning step, the nucleotide analogue 5-methyl-dCTP was added to the nucleotide mixture and [a-32P]dATP was added to monitor the quantity and quality of first strand synthesis. After second-strand synthesis and blunting of cDNA termini, EcoRI adapters were ligated, followed by kinase treatment and digestion with XhoI. The cDNAs were then size-fractionated using Sephacryl S-500 column and then ligated into EcoRI and XhoI digested Lambda Zap Express vector. The ligation product was packaged using Gigapack II packaging extract. The library had primary titre of approx. 1x106. Clones from the primary library were randomly selected for single pass sequencing.
R. Site1:EcoRI
R. Site2:XhoI
Lab Host:
Vector:Lambda Zap Express
Vector Type:Phage
Tissue Supplier:
Library Producer: