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Unexpected bands for negative controls of my PCR on the gel

Unexpected bands for negative controls of my PCR on the gel - PCR - Polymerase Chain Reaction Forum

Unexpected bands for negative controls of my PCR on the gel - 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 03-14-2011, 09:14 AM
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Default Re: Unexpected bands for negative controls of my PCR on the gel

hey all, I also have the exact same problem... can I know the best way to prevent contamination?
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Old 03-16-2011, 01:43 PM
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Default Re: Unexpected bands for negative controls of my PCR on the gel

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Originally Posted by forum123 View Post
hey all, I also have the exact same problem... can I know the best way to prevent contamination?
Just throw away the old reagents, and purchase new ones. Don't waste ur time to find out which reagent was contaminated. Use alcohol to wipe out the surface of laminar flow hood and then UV sterile the hood together with other PCR supplies (but not reagents!!) such as pipet, tips, tubes, even gloves before every PCR trial. At least, these work for me
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Old 05-19-2011, 05:14 AM
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Default Re: Unexpected bands for negative controls of my PCR on the gel

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Originally Posted by butters View Post
is the unwanted band as intense as ur positive control? or very faint but slowly increasing when more pcr is perform? where u load ur DNA?
Hi...the positve control of my PCR showed non specific bands slowly increasing when more PCR i perform.....do you have any idea why this is happening
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Old 05-20-2011, 05:52 AM
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Default Re: Unexpected bands for negative controls of my PCR on the gel

Oh, my its been ages since I come here.

Back to point. Slowly increasing in PCR band intensity mean introduction of more template from the previous reaction aka carry over contamination.

You did mentioned positive control right? What about your negative control? Well, I would "bang" on the part you use the same pipette for loading your positive control template right? A plasmid maybe?
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Old 05-22-2011, 02:35 AM
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Default Re: Unexpected bands for negative controls of my PCR on the gel

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Originally Posted by butters View Post
Oh, my its been ages since I come here.

Back to point. Slowly increasing in PCR band intensity mean introduction of more template from the previous reaction aka carry over contamination.

You did mentioned positive control right? What about your negative control? Well, I would "bang" on the part you use the same pipette for loading your positive control template right? A plasmid maybe?
Thanks for answering me…
The negative control of my PCR doesn't show nonspecific bands on the agarosa gel BUT always appears a band with same size as the target band (164pb)
And yes, I use the same pipette for loading the DNA template
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Old 07-01-2013, 03:09 PM
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Default Re: Unexpected bands for negative controls of my PCR on the gel

Please do tell me about this problem of getting bands in NTC, i have tried different conditions of PCR as well as used new reagents and primers.. but didn't get any results. I am getting same bands in my Positive as well as negative control and these bands are not my target bands.
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