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wrong dntp concentration


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2 replies to this topic

#1 helencollins560

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Posted 07 November 2012 - 05:24 PM

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

#2 Curtis

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Posted 07 November 2012 - 07:17 PM

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).

#3 phage434

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Excellent

Posted 07 November 2012 - 08:21 PM

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




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