I am using the following primers for a PCR
5'-CAC TCG TCC GAG GGC AAA GGA ATA-3'
5'-TCA CCC CAT TAT CAG TTT TGT AGG-3'
Product is 1140 bps
I'm using Invitrogen's Platinum PCR SuperMix:
1 ul each primer
12 ul supermix
2 ul template
I am wondering how to calculate the best annealing temp. In the past we have just used a touchdown program but it is long, and I'm wondering if I get it faster entering my own program. I've done gradients before to determine annealing temps, but I don't want to waste supermix right now because our company is on the thin edge $-wise. Suggestions?
We do DNA extractions with 1X TE