IUBio

How to digest 5' overhang ?

Wolfgang Schechinger Wolfgang.Schechinger at med.uni-tuebingen.de
Mon Mar 22 17:06:57 EST 1999


If you want to remove a defined part from the 5' end, I'd suggest 
designing a suitable primer and isolating the fragment by PCR 
using a high fidelity polymerase (Pwo or Pfu). 

Wolfgang


> Date:          22 Mar 1999 21:20:01 GMT
> From:          kirberg at bii.ch (Jorg Kirberg)
> Reply-to:      kirberg at bii.ch (Jorg Kirberg)
> To:            "bionet.molbio.methds-reagnts mail newsgroup" <bionet-news at dl.ac.uk>
> Subject:       Re: How to digest  5' overhang ?

> In article <36F2ACD1.FF0120BC at leonardo.ls.huji.ac.il>,
> yoramg2 at LEONARDO.LS.HUJI.AC.IL (Yoram Gerchman) wrote:
> 
> > Greetings netters.
> > Dose anyone have a good method for digest  5' overhang ? I tried
> > Mung Bean but it cut into my gene. Will it be better in lower
> > temperature (11C) a-la T4 polymerize?
> 
> With mung bean, make a titration series (several 1/5s) of the enzyme
> and incubate at RT. This worked for me.
> 
> Good luck, jorg
> 
> 
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-----                              
Wolfgang Schechinger   
Pathobiochemistry Dept.      
University of Tuebingen, Germany
email: wgschech at med.uni-tuebingen.de * wwWait: http://www.medizin.uni-tuebingen.de/~wgschech/start.htm
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