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Degenerate PCR Size Limit?


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3 replies to this topic

#1 Johny-Five

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Posted 06 April 2009 - 12:48 PM

Hello all,

From personal experience or otherwise, does anyone know an approximate upper size limit of amplicons using degenerate PCR? The largest fragment I am attempting to amplify is 2.8 kb from using eukaryotic cDNA.

Thanks!

#2 HomeBrew

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Posted 06 April 2009 - 01:33 PM

If the degenerate primers anneal to the template, there is no difference between degenerate PCR and "regular" PCR with regard to template length. You should be able to easily amplify 2.8 kb.

#3 Johny-Five

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Posted 07 April 2009 - 06:26 AM

If the degenerate primers anneal to the template, there is no difference between degenerate PCR and "regular" PCR with regard to template length. You should be able to easily amplify 2.8 kb.


Great... thank you HomeBrew!

#4 HomeBrew

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Posted 07 April 2009 - 07:33 PM

...with regard to template length.


Of course, "...with regard to template length" should have been "...with regard to amplicon length". Good luck!




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