Dissuruption Buffer

dpetersn at unixg.ubc.ca dpetersn at unixg.ubc.ca
Tue Aug 2 13:11:34 EST 1994


In article <31krfv$8fn at mserv1.dl.ac.uk> (David Johnston) daj <daj at nhm.ac.uk> writes:
>From: (David Johnston) daj <daj at nhm.ac.uk>
>Subject: Re: Dissuruption Buffer
>Date: 2 Aug 1994 08:09:51 +0100

>On Mon, 1 Aug 1994 18:38:21 GMT,
>  <szcooley at chip.ucdavis.edu> writes:

>>>Does any one have a recipe for Disruption Buffer used in PCR protocol one 
>>individual E. coli colonies? Thanks, 

>dH2O works fine!

>(add approx 1mm2 of scraped colony to 40 ul, heat 95C for 5 mins, use 1ul 
>as template in a 10ul PCR reaction (use 10/25th of a standard PCR reaction 
>mix). 5ul of this will give a screamingly bright band on an agarose gel 
>for screening purposes.)
>DAJ

Heck I just add some scraped cells with a pipet tip to 5 ul H2O and pipet a 
few times to mix.  Then, add 20 ul of my PCR master mix, oil, and toss in the 
thermocycler (with a preheat of 5 min at 94C) and all goes smoothly.  No 
dilution, no extra steps, just remember to change your tips each time so 
there's no carry-over.

Daniel
"Postdocs may be a dime-a-dozen, but I'd rather have a dozen than a dime."



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