Microwave restriction digests

Viraj Master vam2 at midway.uchicago.edu
Tue Sep 20 17:33:08 EST 1994


In article <Prenom.Nom-190994191534 at agrobacterium.rsvs.ulaval.ca>,
Prenom.Nom at rsvs.ulaval.ca (Prenom Nom) wrote:

> An interesting way of speeding up your restriction digests.  Make up your
> digests as usual in a volume of between 10-50 uL.  Then microwave at high
> power for four times for 10 sec with 2 min intervals at room temperature. 
> Then, run on a standard agarose gel.  I was very surprised that it worked. 
> Apparently, the power of the microwave makes a difference.  We have a 900 W
> oven.
>      What I don't understand is why it works at all.  Most enzymes are
> deactivated at elevated temperatures.  Ten mLs of water is very hot after
> 10 secs in the oven, and imagine a 50 uL reaction using the same time.  Can
> anyone explain what's happening?  Blue

A testimonial:

I'm glad Blue posted this today, because I suddenly developed a good reason
to try it!  Because of 'operator error', I suddenly needed to get a
restriction digest done in a hurry, so I tried this microwave digestion
trick.

I digested 2.5micrograms of plasmid with 20 units of SacI and HindIII in a
volume of 20microliters.  Our lab has a 650W microwave with a rotating
tray.  I heated for 15 seconds maximum power, followed by 2 minutes at room
temp.  I repeated this four times as Blue suggested and then ran on a gel. 
The digestion worked very well.  

Viraj


-- 
Viraj Master
Dept. of Organismal Biology and Anatomy
University of Chicago



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