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How to avoid Nonspecific bands on multiplex PCR?

How to avoid Nonspecific bands on multiplex PCR? - PCR - Polymerase Chain Reaction Forum

How to avoid Nonspecific bands on multiplex PCR? - 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 08-21-2009, 06:41 PM
Pipette Filler
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Default How to avoid Nonspecific bands on multiplex PCR?



Hi everyone,
This is Maddy and working on 4 gene primers multiplex PCR. I am supposed to get 4 bands on my multiplex PCR but turns out to be 6 clear bands on all 53-64 degree Celcius annealing temperatures. I want to get rid of extra two bands that are bigger in size than other bands. My other bands are perfectly on the size range as I expected. I want some suggestions if anybody can answer my questions.
Thank you
TSU
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Old 08-24-2009, 02:32 AM
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Default Re: How to avoid Nonspecific bands on multiplex PCR?

HaHA! I have already seen the great thing!!!
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Old 08-24-2009, 10:13 AM
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Default Re: How to avoid Nonspecific bands on multiplex PCR?

Hi Maddy, try to shorten a little the elongation time in your PCR cycle to prevent the polymerase to amplify longer sequences. Maybe that will help.
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Old 02-24-2010, 09:28 AM
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Default Re: How to avoid Nonspecific bands on multiplex PCR?

Hey anyone ,

Can you answer me to following questions?

1. What is similarities, differences, advantages between nested PCR and Real time PCR?
2. Why is it so difficult to develop a safe vaccine against Dengue fever.Suggestion of strategy to overcome the problem.

I am looking forward for your soonest response.

Thanks a lot in advance,
Alomatxon.
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