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Microwave restriction digests

Koen Dechering koend at sci.kun.nl
Fri Sep 23 09:55:57 EST 1994


In <vam2-200994172455 at beroe.uchicago.edu> vam2 at midway.uchicago.edu (Viraj Master) writes:

>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




The idea is that the number of collisions between the enzyme and its
target are increased by the microwave. It works well for ligations also.


Koen Dechering

A

>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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