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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