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Chlamydia PCR cycling conditions

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#1 icarus.rachel



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Posted 09 March 2010 - 08:25 PM

Hi all,

I'm fresh in running PCR. I have an enquiry about the PCR cycling program on detection genus of Chlamydia.

I'm using the primers stated its Tm is 56-58C.

however, the PCR cycling conditions are as follows

1. 10min at 94C
2. 15sec at 95C for 40cycles
3. 15sec at 55C
4. 30sec at 72C

I have no product for this conditions at the end. I'm thinking to do a gradient to measure the optimum temp for annealing

Please advice for other options. Thank you very much.

Edited by icarus.rachel, 09 March 2010 - 08:32 PM.

#2 leelee



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Posted 10 March 2010 - 05:01 AM

How big is your expected product?

Have you tried various dilutions of your template?

Do your PCR reagents (eg enzyme, dNTPs etc) all work with other reactions?

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