how much dNTP do i add?

Nov 16, 2015
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#1
for my PCR mixture i need final concentration of 432 micromol/ L of each dNTP.



My stock solution from the manufacturers is a dNTP mix of 2.5 mM/800 microliters each dNTP.



the final volume of PCR mic needed is 100 microliters.



can anyone help me i think im doing this all wrong.



Harmony
 

Ruderus

New member
Nov 16, 2015
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#2
Hi Harmony,



take a deep breath.... is a lot less complicate than you think!



1) Your dNTP stock is 2.5 mM (or 2500 µM )



2) You need a final concentration of 432 µM



3) This means that your stock is around 5.8 times more concentrated than it should be in the final reaction mixture (5,8x).



4) If your final reaction volume is 100 ul, and your dNTP have to be diluted 5.8 times, then you need to use:



100 : 5.8 = 17.2 µl of dNTP mix.



The Promega website has useful tools to make all these type of calculations... search for Biomath Calculators. You can also download an App for your smartphone, pretty cool stuff!



Good luck with you PCR