I was wondering if I can use a formula for my ISSR primers to calculate the annealing temperature and melting temperature.
Alternatively I would like to know if is possible to calculate with any existing programme online.
Primer 1 CACACACACACACACAG
Primer 2 GAGAGAGAGAGAGAGAC
Primer 3 CACACACACACACACAT
Primer 4 CTCTCTCTCTCTCTCTRG
I am asking this because I normally use the DreamTaq Kit and with the calculator (on Thermo Scientific) I always find some slight differences compared to the Platinum and Phusion protocol regarding the annealing and melting temperature.
The primers have a reference temperature of:
Annealing °C
Primer 1 47
Primer 2 53
Primer 3 47
Primer 4 48
However, I would like to calculate it if possible.
Thanks in advance for your help.
Best regards