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PCR primers help


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#1 molbio1234

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Posted 29 December 2011 - 04:17 AM

I have a forward primer and a 1:1 mix of 2 types of reverse primers. Both the reverse primers start in the same place, only a few bases are different between them and that they are mixed in 1:1 ratio.

If I use these primers on genomic DNA, will they amplify a product especially since the reverse is a 1:1 mix of 2 different primer sequences? Has anyone heard of others doing this type of approach? Thanks.

#2 phage434

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Posted 29 December 2011 - 06:27 AM

Yes, you will amplify either of the matching sequences.

#3 molbio1234

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Posted 29 December 2011 - 07:05 AM

Phage, thanks for the reply. Do you have any articles that describe this approach? I would like to learn more about it.

#4 hobglobin

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Posted 29 December 2011 - 11:24 AM

google for 'degenerate pcr' or 'wobbles' there are lots of information available.
One must presume that long and short arguments contribute to the same end. - Epicurus
...except casandra's that belong to the funniest, most interesting and imaginative (or over-imaginative?) ones, I suppose.

#5 phage434

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Posted 29 December 2011 - 06:43 PM

You can think of this as a kind of multiplex PCR with primers that just happen to be quite similar.

#6 maverick3006

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Posted 01 January 2012 - 01:24 AM

I use this kind of PCR for determining HLA B27 status of patients. Here, I use two reverse primers to amplify either of two variant of HLA DR gene, which is used as an internal control. The reference is:

Tonks S, Marsh SGE, Bunce M, Bodmer JG. Molecular typing for HLA class I using ARMS-PCR: further developments following the 12th International Histocompatibility Workshop. Tissue Antigens. 1999; 53:175-83.

I hope this helps.




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