RAPID SOUTHERN BLOT PROTOCOL FOR THE ANALYSIS OF TARGETED MUTAGENESIS IN THE MOUSE IGHG1 LOCUS
Keywords:
Ighg1 gene, immunoglobulin, southern blot, embryonic stem cells, gene targetingAbstract
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.
References
Zou Y.R., Muller W., Gu H., Rajewski K. (1994): Cre-loxP-mediated gene
replacement: a mouse strain producing humanized antibodies. Current
Biology, 12, pg. 1099-1103.
Akahori Y, Kurosawa Y. (1997): Nucleotide sequences of all the γ gene
loci of murine immunoglobulin heavy chains. Genomics, vol. 41, Apr., pg.
-104.
Bencsik A. Alena (2008): Cercetări privind obŃinerea şoarecilor de laborator
prin manipulări genetice.Teză de doctorat, USAMVBT Timişoara, pg. 182-