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
>> 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.
>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
>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.
>Dept. of Organismal Biology and Anatomy
>University of Chicago