# PCR mixture - (Aug/24/2008 )

Hi!
I am quite confused:) I am trying to prepare a PCR mixture. In the article which I am folliwing, it says 40umol/L per deoxynucleoside triphosphate and 0.5 umol/L final primer concentration witihn total 10 ul PCR reaction.
for dNTP mixture I couldnt do the convertion of 40 umol/L to mM.Could someone help me for these calculations. (In my lab I have dNTP mixture 2.5mM each)
Thank you all

arfunda

-arfunda-

QUOTE (arfunda @ Aug 24 2008, 12:55 PM)
Hi!
I am quite confused:) I am trying to prepare a PCR mixture. In the article which I am folliwing, it says 40umol/L per deoxynucleoside triphosphate and 0.5 umol/L final primer concentration witihn total 10 ul PCR reaction.
for dNTP mixture I couldnt do the convertion of 40 umol/L to mM.Could someone help me for these calculations. (In my lab I have dNTP mixture 2.5mM each)
Thank you all

arfunda

molarity is the number of moles in a litre of solution, therefore M=mol/L and likewise, umol/L=uM so 40umol/L is the same as 40uM, which is the same as 0.04mM which by the way (in my opinion) seems quite low, I always use 200uM.

-almost a doctor-

Hello
First thank you. Yesterday morning I was just looking the calculations and suddenly I said "Oh my god, I am so crayz they are same, how did I skip this " and this morning I turned on my computer and read your answer. now I am happy that someone else is comfirming me. And yes I agree the amount is quite low but I will try fisrt.
Thank you again
arfunda

QUOTE (almost a doctor @ Aug 25 2008, 01:24 AM)
QUOTE (arfunda @ Aug 24 2008, 12:55 PM)
Hi!
I am quite confused:) I am trying to prepare a PCR mixture. In the article which I am folliwing, it says 40umol/L per deoxynucleoside triphosphate and 0.5 umol/L final primer concentration witihn total 10 ul PCR reaction.
for dNTP mixture I couldnt do the convertion of 40 umol/L to mM.Could someone help me for these calculations. (In my lab I have dNTP mixture 2.5mM each)
Thank you all

arfunda

molarity is the number of moles in a litre of solution, therefore M=mol/L and likewise, umol/L=uM so 40umol/L is the same as 40uM, which is the same as 0.04mM which by the way (in my opinion) seems quite low, I always use 200uM.

-arfunda-