3 replies to this topic

[Johny-Five]

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!

[HomeBrew]

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.

[Johny-Five]

HomeBrew, on Apr 6 2009, 05:33 PM, said:

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!

[HomeBrew]

HomeBrew, on Apr 6 2009, 05:33 PM, said:

...with regard to template length.

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