Do you have any protocol for end-point PCR using meDNA positive and negative control primer pairs in order to see the presence or absence (or very faint band)?
We mainly work with qPCR and not with end-point PCR as the aim is to quantify the enrichment. For end-point PCR, I suggest to run less cycles (25-30) at annealing T 60°C. • Step 1. 95°C 5 min • Step 2. 25-30 cycles of 95°C – 30sec 60°C – 30 sec 72°C -30 sec • Step 3. 72°C 2 min PCR product of 81 bp is expected for positive control and 92 bp for negative control.
