I have small frozen mammalian cell pellets (10-10000 cells) in a few uL of leftover media. I'm planning on doing some qPCR on the DNA using BioRad's iQ Sybr Green. I'm using 2 different sets of primers.
My plan is to resuspend the pellets in 100uL cold H2O and distribute 20uL x 4 wells into the plate wells with my master mixes. Then just hot start the reaction. My thought is that the starting 94C will break open the cells and inactivate DNAses & proteinases enough to start the reaction without first isolating the very tiny amounts of DNA.
Will this work?
Should I be adding something else to help it work?
Thanks for any advice.