purify PCR products using Wizard Prep (Promega)

Ed Stephenson estephen at biology.as.ua.edu
Wed Jun 15 10:24:50 EST 1994


In article <CrE9Gu.Cx7 at acsu.buffalo.edu> vp23qw83 at ubvms.cc.buffalo.edu (Xin Guo) writes:

>Hi, Bionetters:

>I have a trouble to get a high yield of PCR products by using Promega's
>Wizard Prep method. Instead of using lower melting agrose gel, we use
>normal agarose to seperate the PCR products, cut the band, and boil
>it to melt the agarose. 

Having tried some of the "melt agarose and extract the DNA in various ways" 
methods, I have come to the conclusion that none of them is as good as 
elution by electrophoresis into a dialysis bag. There is no kit to buy, no 
phenol extractions to do, it's simple, reliable and fairly quick, and uses a 
piece of equipment that you have anyhow (electrophoresis chamber). We keep a 
supply of 1 cm wide dialysis tubing already cut into 5 cm long pieces in the 
fridge. Place the gel slice and running buffer to fill in the bag and apply 
current until the EtBr-stained DNA is no longer visible in the gel slice 
(monitor with hand-held UV lamp; at 150 volts on our chamber this takes 5 
minutes for a 1-4 kb fragment), then reverse current for 15 seconds to 
dislodge any DNA plastered on the dialysis tubing. Remove buffer with DNA 
and ethanol precipitate.

Ed Stephenson



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