Protocol Online logo
Top : New Forum Archives (2009-): : Molecular Biology

Getting genomic DNA for PCR - (Apr/26/2012 )

Hi,

I wish to know get some genomic DNA (of any kind) for my experiment testing. I happen to saw a JM109 competent cell tube on the freezer. I wonder can I just use that one and run as PCR template?

-deprotection-

If you have E. coli plate, mix with 50ul water, boil for 10 mins, centrifuge high speed for few mins and take supernatant as template.

-christy-

I usually put one colony of E. cloi inside the tube of PCR.
If they are in broth, take as much as a colony and put it inside of PCR tube and mix it with a tip.
It always works.
I have done this more than 50 times without any problem.
We used to call it---> Colony-PCR

Babak

-memari-

Thanks all!

Because I'm not doing cloning (not to mention that I do not have cloning equipment as well), I can't grow them and pick it. I only have the stock amount of competent cells stored in freezer.

Is it possible to take out a few ul of competent cells from the stock and then boils it and spins down for the gDNA as template?

-deprotection-

try that, if you dont have proper facility..... Best wishes.

-christy-