Nicked mitochondiral DNA: how to avoid?

Mike Anson Mike-Anson at NIH.GOV
Sat Jun 17 12:58:12 EST 1995


We've been trying to isolate mitochondrial DNA 
from rat liver. We've tried several protocols, 
but in general, we homogenize the tissue, 
isolate the mitochondria by differential 
centrifugation, lyse with an SDS-containing 
buffer, and purify the DNA in one of the usual 
ways. The details don't seem to change the 
final result: nearly 100% open-circle, no 
supercoiled.

Has anyone encountered this problem, and found 
a solution?

Suggestions would be greatly appreciated!

Thanks in advance.

Mike Anson
Mike-Anson at nih.gov




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