international online community for dna barcoding professionals
I am currently working on DNA Barcoding of some plants (belonging to Angiosperm). Fistly ı did not use DMSO for MatK gene region. But sequence results are not good and then ı use the DMSO, but so now non-spesific band is occurring. Does anyone know why this might be? How can I do ??
Which primers you are using, becoz I am also facing the same problem.
I use primers trnK-2R and trnK3914F ... ;Is it same your primers??
I read in an article that DMSO lowers the annealing temperature of your primers, so if you add DMSO, you will have to adjust your cycling conditions.
I use primers 1248R (Tm 52.5C) and 472F (Tm 57.8C) and my PCR annealing temp is 53.7 when I add DMSO, and I have excellent amplification. Good luck!
I understood, I will try to change my temperature. I hope, I get a good amplification...
thank you so much ..
Good luck, I hope the advice helped!