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How to avoid nonspecific amplification (band) in PCR - (Jul/26/2013 )

How to avoid nonspecific amplification in PCR from PCR?



check the magnesium concentration you used as well as annealing temperature. Usually high mg+ results in non specific amplification


Gradient PCR to set specific Tm for the target gene.


very specific (and long?) primers


higher annealing temperatures help too

-Ameya P-

Several additives such as:

dimethylsulfoxide (DMSO)




tetramethylammonium chloride (TMAC)

polyethylene glycol (PEG)

can help but not necessarily.

Some are also seen as additives that facilitate the reaction, increasing yield or work against inhibitors, the functions seem to differ between authors and are somewhat difficult to define precisely. So it's trial and error, as with the recommended concentrations.