Direct PCR from Gram positive bacteria - Is it possible? (Jul/15/2008 )

Hey guys,

I read this somewhere in this forum (it is gone now) that it is harder to do a direct PCR from gram positive bacteria than gram negative bacteria like E.coli because gram positive bacteria has thicker peptidoglycan.

my question is is it still possible to amplify DNA from the colony itself? I doubt so.... Any input? Thanks!

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Check the first link:
http://search.vadlo.com/b/q?sn=158621799&a...y+PCR&rel=0
..

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thanks cell counter! That was a good idea to break the cells by thermocycler.

I have another question. I have 2 set of primers. And thus this will make it as a multiplex PCR right? Direct pcr from gram positive and multiplex... hmmm do you think it might work?

thanks!

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Well, I never worked with bacterial PCRs, let alone gram troubles

But I do mouse tail lysate pcrs (which have detergents and etc) and quite dirty.

Multiplexing works just fine with even huge mouse genome there, so I would think a simple bacterial genome should not be a problem, as long as you have cracked the bugs.

It finally depends upon the primer compatibility. I would definitely check for 3 prime dimers and etc.

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