I have tried to cut mouse gDNA using the Sall enzyme (TAKARA) but the result was a band at the size of the undigest genomic DNA and a very little smearing downwards. I have added more enzyme in the reaction but the result was the same. Can anyone help with this issue?
SalI activity is blocked with CpG methylation. May be the restriction sites are methylated. Also, what is the percentage of the gel. Smearing may be caused by nuclease contamination or non-ideal digestion conditions. Can you please share the gel pic for better suggestions.
The gel is 0.8%. I have tried different digestion conditions without better result. How can i check if the restriction sites are methylated throughout the whole genome?
Hi,
normally you might rather assume, that the DNA is methylated as this is the predominant state. I agree that this might be the reason here. Would it be possible for you to got with another CpG insensitive enzyme?
Sven
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