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Old 08-14-2017, 03:26 AM   #1
Junior Member
Location: Colombia

Join Date: Aug 2017
Posts: 1
Default Help with PCR

Good morning,

I'm student of master and my thesis is with animals virus. I have problems with the PCR: I ruled out contamination of the reactives, but in the positive samples appear a unknown band, the negative control is negative.

Can somebody help me with papers or handbooks that talk about mistakes in PCR?

Thanks for all.
julyduva is offline   Reply With Quote
Old 08-21-2017, 04:25 AM   #2
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Location: Munich

Join Date: Jun 2017
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Can you give us a little more information, it may be that the unknown band is due to non-specific binding of your primers? What size is it roughly? What cycling conditions did you use, composition of your master mix etc? Bio-rad PCR troubleshooting is always a good place to start:
asorbie is offline   Reply With Quote
Old 08-21-2017, 10:39 PM   #3
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Location: The Netherlands

Join Date: May 2017
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In your positive samples, do you see your expected band AND an unknown band or you just see unknown/unexpected band? Is it PCR that is known to produce good results or you are trying to develop a PCR ?
Dorota is offline   Reply With Quote

pcr artifacts, pcr errors

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