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Bands in PCR negative control

Started by Fhannan, May 19 2010 06:34 AM

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3 replies to this topic

#1 Fhannan

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Posted 19 May 2010 - 06:34 AM

Hello, i am new here and i wish if anybody can help....
I came across a problem recently is that the negative controls of my PCR showed a band on the agarose gel. The weird thing is that it has the same band size as my target band 305bp. What might be the cause? Do I have to repeat the samples which have been analyzed without running a neg control along with them as they might be just contaminants and not the real targeted DNA sample?

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#2 Trof

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Posted 19 May 2010 - 06:37 AM

Try things suggested in this topic.
http://www.protocol-...showtopic=14306

Our country has a serious deficiency in lighthouses. I assume the main reason is that we have no sea.

I never trust anything that can't be doubted.

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#3 ksam

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Posted 27 May 2010 - 11:42 AM

I think the only reason for getting band in negative control is primer contamination. make new stock of primers and do PCR. Good luck!

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#4 bob1

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Posted 27 May 2010 - 04:15 PM

ksam, on May 27 2010, 12:42 PM, said:

I think the only reason for getting band in negative control is primer contamination. make new stock of primers and do PCR. Good luck!

Contamination could be in any of the reagents.

Quote

Hello, i am new here and i wish if anybody can help....
I came across a problem recently is that the negative controls of my PCR showed a band on the agarose gel. The weird thing is that it has the same band size as my target band 305bp. What might be the cause? Do I have to repeat the samples which have been analyzed without running a neg control along with them as they might be just contaminants and not the real targeted DNA sample?

The contamination is probably a PCR product that has somehow made its way into your reagents (possibly from your pipettes). You do need to repeat the samples from that PCR as they may well be false positives.