High-throughput semi-quantitative analysis of insertional mutations in heterogeneous tumors

ABSTRACT: Exploiting the full potential of insertional mutagenesis screens with retroviruses and transposons requires methods for distinguishing clonal from subclonal insertion events within heterogeneous tumor cell populations. Current protocols, based on ligation mediated PCR, depend on endonuclease based fragmentation of genomic DNA, resulting in strong biases in amplification and sequencing due to a fixed product sizes of the amplicon. We have developed a method called shear-splink, which enables the semi-quantitative high-throughput sequence analysis of insertional mutations, enabling us to count the number of cells harboring a given integration, within a heterogeneous sample. The shear-splink method enriches for (sub)clonal integrations, thereby reducing the contribution of irrelevant passenge

ORGANISM(S): Mus musculus

SUBMITTER: Jos Jonkers 

PROVIDER: E-GEOD-22657 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress