I had a problem about multiplex-PCR - (Aug/29/2001 )
i had a problem about multiplex PCR. my result showed nonspecific bands . My work had five primers in one single tube and amplified for only one band but it show non specific bands, so I need to knew additive or optimized condition for PCR for specific productoh, my primers have varies Tm. Now I use annealing temperature is 65 C Thanks for answer.
I suggest that you use the sample preparation strategy. The problem is substances in your sample that affect theamplification. You need to purify your sample prior to PCR. What is your sample?
I use culture and serum for test.I try vary Mg2+ , pH , and concentration of primer ,and annealing temp. I found that vary concentration primer has good response but not very good result because it has non specific band .Help me!!!!!!!!!!!please.......
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