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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.