PCR re-amplification

Yves Bertheau bertheau at inapv.inapg.inra.fr
Thu Jun 4 12:14:05 EST 1992


In article <1992Jun2.235739.7982 at usenet.ins.cwru.edu> axa12 at po.CWRU.Edu (Ashok Aiyar) writes:
>
>In a previous article, listona at bionette.cgrb.orst.edu (Aaron Liston) says:
>
>>I would like to hear from anyone who is successfully re-amplifying
>>PCR products.  How do you do it?
>>Thanks.
>
>Gel purified PCR product (electro-elution onto DE81 paper or
>DEAE Nylon membrane) works fine as a template for re-amplification.
>I've re-amplified products from 300 bp to about 2.5 Kb.  
>
>Whenever I've tried the re-amplification without purifying the
>product, I've ended up with tons of primer-dimer.
>
>
>Ashok
>-- 
>  Ashok A. Aiyar   Department of Biochemistry   CWRU Med. School
>   		       axa12 at po.cwru.edu
>		   aiyar at cwbio.bioc.cwru.edu
>                Visit the IBM-PC Sig on Freenet

All the answers I am reading are concerned by a new amplification after
purification of the amplified fragment(s).

But did somebody a re-amplification directly in the "first" mixture or from
an aliquot, e.g.  because the amplification is very slight, or for practical
purposes (as routine diagnosis) ?

In this case is the "nested" PCR the only way to re-amplify ?

Thanks in advance
-- 
		Yves Bertheau
INRA INA P-G		Pathologie Vegetale	16 rue Claude Bernard	
75231 PARIS cedex 05	FRANCE	Tel +33 (1) 43.31.93.97 
Fax: +33 (1) 43.31.83.82 Internet: bertheau at inapg.inra.fr



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