I am using your CpG WIZ MGMT amplification kit for gliomas cases. Currently I am standardizing the assay and I have followed the instructions mentioned in the kit insert. I am using Qiagen DNA kit for extraction of DNA from FFPE tissue . I used 15 ul (approx 1 ug) of the FFPE DNA for bisulfite conversion using Epitect kit from Qiagen and did the PCR amplification . The gel pic post PCR amplification shows that except for the control DNA (universal methylated DNA) the samples did not showed any amplification with very very fain amplification with unmethylated primers while with methylated primers no amplifcation was observed. Can you please suggest me how to improve the amplification intensity of the amplicons?
The input DNA samples looked of without much degradation and I feel that it is unlikely that the DNA is degraded upto that extent that it cannot under go chemical modification. Even the modification was done one day prior to PCR.
Looking forward to hear from you at the earliest.
Thank you all in advance for the suggestions