The final concentration of primer that is usually included in a PCR mix is 1 - 5

Question

The final concentration of primer that is usually included in a PCR mix is 1 - 5 uM. This often requires preparing a more dilute stock solution before adding primer to the PCR mix. Describe how you would prepare a 25 uM solution of your oligonucleotide (100 ul final volume) using a 1 mM stock solution of the primer. Show your work

Calculate the amount of MgCl2 you would need to make a 50 ml solution of 125 mM MgCl2 (show your workand units and circle your answer ) Molecular weight of MgCl2 is 95.2 g/mol

for the first question, you are using c1v1=c2v2 equation to answer for how you would prepare a 25 um using 1mm stock solution

Explanation / Answer

1) 1 mM = 10^3 micro Molar.

So applying the formulae

C1V1(stock) = C2V2(solution to be prepared)

10^3 * V1 = 25* 100

V1 = 2500/1000

V1= 2.5 microlitre..!!

2) applying the formulae of molarity

M = (Given mass * 1000)/ (Molecular mass * volume of solutions)

1 M = 10^3 mM.. So it is 0.125 M..

0.125 = (Given mass *1000)/ (95.2 * 50)

Given mass = (0.125*95.2*50)/ 1000

=> 0.595 gm

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