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Getting genomic DNA for PCR


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4 replies to this topic

#1 deprotection

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Posted 25 April 2012 - 11:24 PM

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?

#2 christy

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Posted 26 April 2012 - 12:36 AM

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.

#3 memari

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Posted 26 April 2012 - 05:38 PM

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

Edited by memari, 26 April 2012 - 05:40 PM.

-----
Babak Memari

#4 deprotection

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Posted 26 April 2012 - 07:52 PM

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?

#5 christy

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Posted 26 April 2012 - 08:04 PM

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




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