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PCR for the 1ST TIME

PCR for the 1ST TIME - PCR - Polymerase Chain Reaction Forum

PCR for the 1ST TIME - 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 08-15-2011, 09:16 AM
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Wink PCR for the 1ST TIME



I have been trying to do my first PCR but met with failures.

The 1st PCR showed 1kb bands in the agarose gel lane of my water control, pcDNA3.1 vector control, and the plasmid DNA template. I am trying to amplify DC-SIGN gene of 1200bp. I have been using plasmid DNA concn of 50ng/ul.

I did another one that showed no bands at all.

I then repeated the PCR using Supertherm Taq Polymerase instead of Phusion Taq polymerase that I used in previous reactions.

What could be wrong with my procedures?

Thanks a lot.

Yusuf
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Old 08-20-2011, 04:59 AM
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Default Re: PCR for the 1ST TIME

It sounds like cross contamination of PCR product, perhaps during PCR preparation. Must ensure the water for PCR is clean without any DNA. Additionally, different Taq polymerase will have some slight difference in the denaturation temperature or extension temperature, you should check on the manufacturer manual.
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