I have these two primers F: TCAAAGGAATGTTGGG and R: ACGCACATAGGGATTG to use in PCR. I want to know what is the Tm for the two primers in order to know what annealing temperature to be used while running the PCR.
I will be using this buffer 10X ViBuffer A (without MgCl2} 500mM KCl, 100mM Tris-HCI (PH 9.1 at 20°C) and 0.1 % Triton™ X-100. The buffer is optimized for use with 0.1-0.2mM of each dNTP. where the primers concentration is 2um.and the MgCl2 is 50mM.
Please I need some help with these two primers F: TCAAAGGAATGTTGGG and R: ACGCACATAGGGATTG . Any help is really appreciated I have calculate Tm using Wallace rule
Tm = 4(G+C) + 2(A+T) Tm Forward primer = 46 C and for Reverse =50 C. In addition to that I used online Tm calculator at
thank you very much,
Please need some help