High-throughput semiquantitative analysis of insertional mutations in heterogeneous tumors.

ABSTRACT: Retroviral and transposon-based insertional mutagenesis (IM) screens are widely used for cancer gene discovery in mice. Exploiting the full potential of IM screens requires methods for high-throughput sequencing and mapping of transposon and retroviral insertion sites. Current protocols are based on ligation-mediated PCR amplification of junction fragments from restriction endonuclease-digested genomic DNA, resulting in amplification biases due to uneven genomic distribution of restriction enzyme recognition sites. Consequently, sequence coverage cannot be used to assess the clonality of individual insertions. We have developed a novel method, called shear-splink, for the semiquantitative high-throughput analysis of insertional mutations. Shear-splink employs random fragmentation of genomi

SUBMITTER: Koudijs MJ 

PROVIDER: S-EPMC3227106 | biostudies-literature | 2011 Dec

REPOSITORIES: biostudies-literature