2 replies to this topic

[helencollins560]

Hi

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

[Curtis]

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).

[phage434]

You may need slightly higher magnesium concentration when using larger dNTP concentrations.