wrong dntp concentration - (Nov/07/2012 )
just performed and RT and following real-time pcr using 10mM dntp instead of 2.5mM dNTP for both RT and realtime
just wondered whether i would get any usable data from that? or whether as my gut tells me that i will need to complete the plates again
any help, advice would be much appreciated
It should be OK. We add 1-2.5 ul dNTP (10mM) to 50 ul reaction. I have seen protocols that use even more (KOD polymerase for example).
You may need slightly higher magnesium concentration when using larger dNTP concentrations.