Any one can help me to find out
the annealing temperature to this sequence
β-Lactoglobulin Gene 1 ((452bp exon II ))
forward 5’-AAA AGC CCT GGG TGG GCA GC-3’
reverse 5’-TTG GGT TCA GTG TGA GTC TGG -3’
I try different way to get the PCR product from this sequence but
i don't get any thing.
Paul Rutland
the melting temperatures of these primers are 64 and 57c so I would run a gradient of annealing temperatures of 52-60 and see what works. Are you getting a primer dimer or any other wrong size bands?. Can we see a picture of the checker gel and pcr conditions please?. Have you checked the sequences using BLAST or primer blast to make sure the sequences are correct and that there are no polymorphisms under the primer annealing positions?
2 votes
1 thanks
- Badges
- Science method
More Ahmed A. Mnati's questions See All
Similar questions and discussions