DNA from gel and questions

frandenm at TCPLINK.NREL.GOV frandenm at TCPLINK.NREL.GOV
Thu Jul 14 11:47:04 EST 1994


     
     I use this same method and although I've never quantified it, it 
     seems recoveries are decent.  If you use TAE, you can dissolve gel 
     slice in 3X vol of NaI/wt of gel slice.  If you use TBE, you can use 
     TBE modifier soln which comes in Gene Clean kit from BIO101.  Then 
     incubate at 55oC for approx. 10 min.  
     
     Mary Ann Franden
     
______________________________ Reply Separator _________________________________
Subject: DNA from gel and questions
Author:  dxy at iastate.edu (Dongxian Yue) at smtp 
Date:    7/14/94 12:09 AM
     
     
Dear frinends,
     
Too many ways to get DNA from agarose gel. Here is another one. 
     
You dissolve the gel slice in NaI, then mix with Promega Wizard Resin, 
follow the Wizard procedure, your DNA will be in H2O in minutes.
     
So there is no ethanol ppt. 
     
No elution, no mess up. 
     
No phenol/chloroform.
     
Works well for me.
     
Here is the questions: When agarose is dissolved in NaI, what happened 
to the agarose? Is it cleaved or what? 
     
Is there solvent could dissolve polyacrylamide gel and do not harm RNA? 
I need to get small RNA from denature gel, all the methods are so nesty.
     
A question for Amersham mutangenesis kit users: This kit uses 
[alpha]S-dCTP to synthesize the mutant strand. Can this strand survive 
in vivo? 
     
Welcome comments.
     
Dongxian Yue
     
-- 
     
     



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