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shifali chatrath via methods%40net.bio.net (by shifalich from rediffmail.com)
Wed Nov 26 20:39:36 EST 2008


Dear Gayacharan!
The amplification that you will be obtaining like this will be geometric and not exponential. That, as may number of cycles, as is the number fold the amplification will be. That means, you cannot use the number of cycles as is used in normal PCR. Also, check the polarity (5' or 3')of ur primer as well as template, if you know primer designing. Also, in order to get amplification of your ss DNA,if you use too many cycles, I doubt the fidelity of the reaction. So, all these factors you need to take into account.

All the best for your experiment.

Shifali


On Thu, 27 Nov 2008 04:04:33 +0530  wrote
>I want to amplify single strand DNA by a single gene specific primer from genomic DNA. I used thermal cycler for this purpose, but I am unable to get the result. I want suggestions regarding this.
>   
>  Thank you
>   
>  Birendra, P.
>
>
>GAYACHARAN
>M.Sc. Biotechnology
>Lab - 210, Rice Stress Molecular Laboratory
>Centre for Plant Molecular Biology
>Tamil Nadu Agri. Uinversity,
>Coimbatore- 641003
>INDIA
>       
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Shifali Chatrath
Graduate Student
Protein science Lab
Dept. of Biological sciences
National University of Singapore
Singapore
+65-65161210
+65-96393449


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