purify PCR products using Wizard Prep (Promega)
Sean David Moore
smoore at mail.sas.upenn.edu
Tue Jun 14 15:41:59 EST 1994
Xin Guo (vp23qw83 at ubvms.cc.buffalo.edu) wrote:
: 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. Then I follow the Promega's steps to mix the
: DNA products with the resin, add it into the column, and elute it.
: It is said the recovery can be >90%, but we only get <10%. Are there
: any of you has ever utilized this method to purify DNA from agarose gel?
: Any suggestion will be great appreciated.
I think the problem is the fact you are using TBE as buffer...borate seems
to screw with almost all of the "resin binding" kits.
If you have a good band, just clean it as is...if you have multiples...
use TEAE or similar buffer and low-melt the gel...the resin will work fine...
smoore at sas.upenn.edu
baldy at brahms.udel.edu
TATABOX at aol.com
: Xin Guo
: SUNY at Buffalo
: Department of Pharmacology and Toxicology
: vp23qw83 at ubvms.cc.buffalo.edu
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