How to perform colony PCR - (Mar/02/2013 )

Hi,

I am new to the PCR. Now I dealing with the colony PCR, I have a problem which prevails like a stubborn thing. I working with gram negative bacteria which lacks in peptidoglycan and it is easy to extract the DNA from those. My problem is, I tried the template preparation by the usual method. But, I getting the template DNA sometimes luckily and for the most of times it isn’t working. I tried the other methods of template preparation by using lysis buffer. Even though the problem persists.

Can anyone solve this issue?

Thanks in advance

we already have these guidelines at protocol-online.org but this video might also be helpful. I follow this method too.
http://www.youtube.com/watch?v=h0yRrDWtdA4