how much DNA you need for real time pcr after chip
Posted 05 September 2009 - 08:50 AM
after ChIP i have resuspended the pellet in 40 ul of Tris, 10mM and i got a concentration of 2-4 ng/ul.
is that enough to run a real time pcr?
How much do u use?
Posted 05 September 2009 - 01:28 PM
in theory, for realtime PCR a Ct of 40 is required to amplify one copy of the template of interest. The fact that you have DNA there should be fine.
for methylation enrichment experiements (similar to ChIP) I would use 0.5-1ul of sample (without measuring it on nanodrop) because I have a fair idea it would be very low in concentration and don't want to waste it anyway.
All comments and communication are my own personal ones, and are not tied to any of my affiliations.