RAPID SOUTHERN BLOT PROTOCOL FOR THE ANALYSIS OF TARGETED MUTAGENESIS IN THE MOUSE IGHG1 LOCUS

Abstract

The efficiency of homologous recombination relies on the lenght of the homology arms and the chromatin compactation of the locus. In ES cells the immuoglobulin locu is compacted. Thus, the homologous recombination events occur at a very low frequency. A high number of ES cell colonies have to be analysed to detect a homologous recombination event. Here we describe a reliable and rapid protocol for the analysis of targeted mutagenesis in the immunoglobulin locus using Southern Blot analysis.

Keywords

• ighg1 gene
• immunoglobulin
• southern blot
• embryonic stem cells
• gene targeting