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| I even use primer design software, what results Self Dimer dG Mb u need to order/choose another primers? Another way is: - Set anneling time to minimun - set concentration of primers to minimum - increase Ta or if u detect dimers only in low concentrations of substract u can try to add incubation b4 Plate Read just around 1-2 sec (Temperature: hi then T denaturation of dimers, but lower then T denaturation of ur product) This information was taken form molbiol.ru Sorry for my bad english ![]() |
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| I first suggest that the primer concentration be set to a minimum after adjusting for optimal primer:target sequence ratio. I also suggest that one need to re-design the primers such that the last three NTDs at 3' end contain no G or Cs. Increasing the annealing temperature will not help much if the primer:target ratio is far from optimum |
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