Hello,
If any body have verified protocol for extraction of Mega Plasmids from Bacteria, please share.
I have tried to extract the Mega Plasmid from Gram negative bacteria but could not able to resolve on Agarose gel (0.6%). I tried with SDS also. There is something which was trap in the well but did not moved. Please share your working protocol.
Attached are some "old" papers with probed protocols for megaplasmid extraction.
But, you said that you're not able to resolve in 0.6% agarose. I suggest to follow the exact electrophoresis conditions of paper of Anderson etal.. The band that you're seeing at the wells is damaged megaplasmid, that is unable to run in simple unidirectional agarose gel. You could be able to force these samples to run in a PFGE, but you need all the instrumental.
hope this helps
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