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how much DNA you need for real time pcr after chip


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#1 sushina

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Posted 05 September 2009 - 08:50 AM

Dear all,
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?
Thanks!

#2 methylnick

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Posted 05 September 2009 - 01:28 PM

hi sushiina,

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.

cheers

Nick




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