PCR primers

Errol E.S.Kwan at massey.ac.nz
Sun Jul 30 19:34:08 EST 2000


I think the primers should be at the other ends of the DNA and facing
the opposite direction.  Primer 1 should be at the 3' end of the top
strand and facing the other way, so the primer has it's 5' end at the
DNA stands 3' end and the extension will go from the 3' end of the
primer.  Likewise the Primer 2.

On Tue, 25 Jul 2000 13:07:08 +0100, "Dr. Duncan Clark"
<drc at nospam.demon.co.uk> wrote:

>In article <%v4f5.1081$WM6.81753626 at news1.mtl.metronet.ca>, Tina
><anfortin at webnet.qc.ca> writes
>>It is possible to find the location of upper and  lower primers in the gene
>>sequence in order to know if or not they will generate a PCR product
>>included in one exon or covering two exons.
>>For the upper primer, we have just to find it directly in the gene sequence,
>>but for the lower primer it is necessary to "translate" the primer sequence
>>in complementary bases and after to invert it before searching it in the
>>gene sequence. Why the upper ca be find directly and not the lower ?
>>I can't stay more time without undertanding this basic point of PCR
>>technique .
>
>Think DNA not gene sequence. DNA is double stranded and the product you
>amplify will also be double stranded, therefore you need primers
>complementary to both strands i.e. an upper and lower primer, one for
>each strand.
>
> Primer 1
> -------->
>5------------------------------------------------3
>3------------------------------------------------5
>                                        <--------
>                                         Primer 2
>
>Duncan
>-- 
>The problem with being on the cutting edge is that you occasionally get 
>sliced from time to time....
>
>Duncan Clark
>DNAmp Ltd.
>Tel: +44(0)1252376288
>FAX: +44(0)8701640382
>http://www.dnamp.com
>http://www.genesys.demon.co.uk






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