PCR problem

divya venugopal divyavenugopal at hotmail.com
Tue Feb 6 12:08:13 EST 2001

Can anybody help me out with this:

I'm trying to amplify a 1.5kb region from the origin of replication of a 
plasmid from Mycobacterium avium-intracellulare.  The primers have been 
designed from the published sequence of the origin of replication.  The 
calculated annealing temperature of the primers is 67 degree C.  However i 
did not get any results at that temperature.  I got an amplicon at 55 degree 
C.  The size works out to be 0.5kb.  What could this be as i should be 
getting a 1.5 kb band. How is it possible to get the expected band.

thank you,


>From: Tim Spahlinger <txs at po.cwru.edu>
>To: methods at hgmp.mrc.ac.uk
>Subject: Re: primers problem
>Date: Tue, 06 Feb 2001 10:42:48 -0500
>Tina wrote:
>>Can we dilute primers in some buffer instead of water. If yes, which 
>>Is there a special task with porcine primers?
>>Does water have any effect on the primers?
>>Any suggestions ?
>>Thanks a lot !
>I've used TE buffer to reconstitute and freeze (-20 C)
>commercially prepared primers.

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