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how to dilute 100mM dNTP to 1.25mM? what is the calculation to make pcr reaction...

how to dilute 100mM dNTP to 1.25mM? what is the calculation to make pcr reaction... - PCR - Polymerase Chain Reaction Forum

how to dilute 100mM dNTP to 1.25mM? what is the calculation to make pcr reaction... - PCR - Polymerase Chain Reaction Forum. Discuss and ask questions about PCR troubleshooting, PCR protocols and methods, PCR products, and PCR theory.


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Old 09-21-2011, 11:16 PM
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Default how to dilute 100mM dNTP to 1.25mM? what is the calculation to make pcr reaction...



...work solution? I have stock of 100mM dNTP. I need to make 1.25mM for 25ul pcr reaction.I am confused about the calculation.If anybody help me please...
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Old 09-22-2011, 05:50 AM
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Default Re: how to dilute 100mM dNTP to 1.25mM? what is the calculation to make pcr reaction.

hola, 100/1.25=80 so you have to dilute your solution 80 times to have 1.25mM. But if this concentration is the final in reaction mixture, 25/80=0.3125ul 0r 3.12 ul of an intermediate solution 10mM (10ul of 100mM +90 H2O). Buena suerte
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Old 09-23-2011, 09:18 AM
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Default Re: how to dilute 100mM dNTP to 1.25mM? what is the calculation to make pcr reaction.

The best and the most used formula I ever learned at school is this one:
c1*v1=c2*v2 where:
c1= the concentration of the solution you want to dilute
v2= the volume you should use of the solution
c2= the concentration you want in the final solution
v2= The volume you want of the final solution

So lets say you want to make 500 ul of a 12,5 mM solution the formula is:
c1*v1=c2*v2
100 mM * v1 = 12,5 mM * 500 ul
v1 = 62,5 ul
That means you have to use 62,5 ul of your 100 mM solution and dilute it in 437,5 ul of whatever you want to dilute it in to get a final volume of 500 ul. Then you have a 12,5 mM solution of your primers, and you can then use 2,5 ul of this solution in your 25 ul to get a final concentration in the reaction of 1,25 mM.
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Old 12-10-2011, 06:23 AM
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Default Re: how to dilute 100mM dNTP to 1.25mM? what is the calculation to make pcr reaction.

Very good answer
mahmoud manama
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