has anyone here ever amplified chloroplast matK gene before? - (Apr/28/2008 )
Has anyone here ever amplified chloroplast matK gene before? I found the univeral primers in some articles and the sequences are as follows:
MG1: 5' CTA CTG CAG AAC TAG TCG GAT GGA GTA GAT Tm=71 degrees
MG15: 5' ATC TGG GTT GCT AAC TCA ATG Tm=57 degrees
However, I found there are large melting temperature differences between this two primers. How they will work in PCR as the paper said the working temperature for annealing is 64 centi degrees?
I would suggest you just try it, published primers and conditions usually work!
you could do gradient and see which temperature works best.