Plasmid methylation problem - Plasmid methylation (May/20/2005 )

Hi, everyone. I'm looking for a protocol to strip methyl groups from a plasmid. I'm currently trying to digest a plasmid with XbaI and I'm having difficulties since XbaI sites are methylation sensitive. Does anyone know how I can get around this problem? Thanks!

Use an isoschizimer of XbaI, try looking at REBase http://rebase.neb.com for one.

Just looked, there isn't an isochizimer so you will have to transform your plasmid into a methylation negative bacteria cell like DH5 or something along those lines.

Good luck!

nick

I also have a problem with Xba I digestion. My plasmid was extracted from DH5 cells. I have also sequenced my plasmid and know that there is a Xba I site. Could there other problems besides methylation?