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PCR Smear

PCR Smear - PCR - Polymerase Chain Reaction Forum

PCR Smear - 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 04-21-2011, 10:43 AM
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Default PCR Smear



hi
i am amplifying a 2kb gene but geting a smear with the product.Changed water other components, diluted dna,different magnesium conc. but nothing worked. Smear still presist with the product (2kb).
thanx
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Old 04-21-2011, 10:50 AM
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Default Re: PCR Smear

Hi welcome!
Have you ran your template alone to see if it is the cause of the smearing?

Is your PCR freshly run on gel? It could be degraded. Also can you try to purify your template before you PCR to get rid of any DNase?

Have you tried to run on a different gel or checked to see if your ladder is running clearly to see if the gel running is an issue?
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