Does dNTP bottle concentration mean total or per nucleotide?

[sotellme]

Does the concentration of dNTP in the bottle's label specified the total concentration of the four nucleotides or only each of them? When it says in the protocol "each 200uM" does it mean that i have to add a concentration 4 fold (200uM * 4) of the bottle to get all the four different nucleotides or do i only need a final concentration of 200uM in the PCR reaction? The label says 5mM.

 

[Moonbear]

You can double check with the manufacturer (always a good idea when unsure about product labeling), but it should be the concentration of each dNTP.

 

[iansmith]

The protocol states that you need 200 uM of each nucleotides, then you need to reach to concentration for each nucleotides if they come in separate tubes. Some compagnies have set of 4 tubes for each nucleotides and also have a premixed nucleotides solution.
If you have premixed dNTPs then you just add a certain volume of the dTNPs mix to reach 200 uM. Each nucleotides will have a concentration of 200 uM. So for a 50 uL PCR you will need 2 uL of your 5 mM dNTP mix.