STANDARDISATION OF MULTIPLEX PCR - IN order to standardise a multiplex PCR for 3 set of primers (May/26/2011 )
we like to standardise a multiplex PCR for BACTERIA, P.acne, Fungal from a clinical samples. when I did Multiplex PCR am getting only fungal and p.acne POSITIVE but negative for bacteria, but when the same condition and reaction in UNIPLEX it gives good result for all. what i should do now???
and i like to know whether 15mM Mgcl2 which is am using it.But literature says for that use 5mM Mgcl2. Whether this may inhibit the multiplex or not??
waiting for your valuable suggestion.
hmn.. had the paper been published before? you design the primers your own?
Adrian K on Thu May 26 21:12:44 2011 said:
hmn.. had the paper been published before? you design the primers your own?
yes sir, its already published paper only. i just followed all as such in paper.
okie, did you perform a gradient PCR? this is because temperature varies from machine to machine... although it may published 55C...but you might need push up further to 58C...
can you show us the paper? any links?