I am using EvaGreen (like SYBA Greeen) run qPCR, and got GAPDH non specific product that has higher melting temp on dissociation curve.
Tm of target product is 81C, the non specific product is 87C
Maybe the primer is too short (68bp), this primer pair passed primer dimer and second structure check.
Could you please tell me your thoughts
Non specific product has higher melting Temp than target
No replies to this topic