Bacteriophage genome extraction - (Jan/17/2013 )
We have recently a bacteriophage which appears to be a Siphoviridae. It has got a small genome of about 20 Kbp. I isolated the genome with standard phenol/ chloroform extraction and ethanol precipitation but after extraction and when I want to digest, the genome seems to be degraded and smear on the gel. I checked all the reagents used in the experiment but they all seem to be fine and work with other + controls. Even if I incubate the extracted DNA in water at 37 C it smears on the gel and if I add a restriction enzyme buffer without the RE and incubate at 37 the DNA gets completely degraded and nothing is visible on the gel. I was just wondering if anybody knows why this is happening and maybe have had similar experience and how to resolve the issue.
Did you redissolve the DNA after precipitation in TE? Resuspension in water can lead to rapid degradation.