pcr from glycerol stock!

Michael Witty mw132 at mole.bio.cam.ac.uk
Thu Dec 6 15:10:56 EST 2001


Dear Sushma,
           I have done this successfully by using a tip and scraping a
little slush off the top of my glycerol stock (0.5ml E. coli culture
0.5ml 80% glycerol) and putting about 1 microlitre in a standard reaction.
Wolfgang is right though - you should try several dilutions to see what
is best for _you_.  Actually I am sometimes surprised by the gunge I can
put into a PCR at still get OK results.  I am ever more surprised when
I put beautifully pure DNA into a PCR and get nothing!
                                                     Regards, Mike.

 On Thu, 6 Dec 2001,
Sushma Tiwari wrote:

> Hi,
> I would like to know if any of you have done PCR directly from a culture
> in glycerol stock. Is it possible, If yes could you share your
> protocols/experiences with me please.
> I have to amplify clones which are in 384 well format (1500 colonies)
> and they are in glycerol stocks, I would have to spend a lot of money
> and energy if i have to grow them individually and padmid prep all of
> them.
> Any ideas will be welcome,
>
> sumis
>
>




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