Find content
Not Telling
hi,
I am going to order my primers from SIGMA, it's need to be desalt primer?
if i used bisulfited treated DNA and use the above primers to run two parallel PCR with the following cycling protocol (I will use the EpiTect MSP from QIAGEN)
initial activation step: 10 min 95c
3-step cycling
denaturing 15s 94c
Annealing 30s 50-55c
Extension 30s 72c
44cycles
final extension 10 min 72c
My reaction composition will be:
25ul Eptic master mix(contain the hot start Taq, d-tec polymerase,Tris.cl, KCl (NH4)SO4, MgCl2 and dNTPs), I ul from each primer ( methylated and Unmethyalted), 1 ul DNA template (bisulfited) and 23ul RNase-free water
Do u think this make sense? and can work and give visible band?
Thanks for ur great help,
Laila
