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Repeatability PCR

Repeatability PCR - PCR - Polymerase Chain Reaction Forum

Repeatability PCR - PCR - Polymerase Chain Reaction Forum. Discuss and ask questions about PCR troubleshooting, PCR protocols and methods, PCR products, and PCR theory.


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Old 03-08-2012, 03:17 PM
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Default Repeatability PCR



Hello everyone,

I have troubles with the repeatability of my results on PCR. Both intensity and presence of the bands are everytime different. First I thought I made some mistakes preparing my master mix, so I did the same PCR's over and over again without any clear result... also my colleages appear to have some difficulties with the same primers and samples I use. Interestingly, till now it's only with specific primers of one specific gene that give an inconsequent result.

Has anyone an idea what is going wrong?

Thanks!
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Old 04-01-2012, 05:03 AM
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Default Re: Repeatability PCR

first , u can check out ur DNA template for degradation
also try new aliqouts of ur primers and use filter tips to avoid contamination
change the electrophoresis running buffer
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Old 01-23-2013, 07:11 AM
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Default Re: Repeatability PCR

[Only registered users see links. ]There are two inputs from my side.First there was variation in the volume of PCR reaction mix after the program ended, even when heated lid was used. Thus I started adding mineral oil and tested all the wells for the same cDNA and primer set. all the wells gave similar results
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