Rapid and quantitative detection of homologous and non-homologous recombination events using three oligonucleotide MLPA.

ABSTRACT: Embryonic stem (ES) cell technology allows modification of the mouse germline from large deletions and insertions to single nucleotide substitutions by homologous recombination. Identification of these rare events demands an accurate and fast detection method. Current methods for detection rely on Southern blotting and/or conventional PCR. Both the techniques have major drawbacks, Southern blotting is time-consuming and PCR can generate false positives. As an alternative, we here demonstrate a novel approach of Multiplex Ligation-dependent Probe Amplification (MLPA) as a quick, quantitative and reliable method for the detection of homologous, non-homologous and incomplete recombination events in ES cell clones. We have adapted MLPA to detect homologous recombinants in ES cell clones target

SUBMITTER: Langerak P 

PROVIDER: S-EPMC1310898 | biostudies-literature | 2005 Dec

REPOSITORIES: biostudies-literature