PRIMER DILUTION - STOCK SOLUTION TO A WORKING STANDARD (Jun/28/2011 )

HI TO EVERY ONE...

THE QUERY WHICH AM POSTING HERE WOULD HELP EVERYONE, WHO DEALING WITH PCR PRIMER--

THE QUERY IS ;;;


How to dilute a STOCK solution of original primer 100microMolar concentration of 1ML..(SIGMA PRIMER)

to

a> 0.36 micromolar
b> 1 micromolar
c> 10 picomoles


in a 50 microlitre PCR mix..

kindly brief the protocol and the principle behind it,,,,



I will be so grateful to all who rectify my query..

Thanks
Regards

Thiru Murugan. N

Dear Ms Thiru Murugan,

You can use the M1V1=M2V2 formula.
M1 = Molarity of the stock solution (100uM)
M2 = Molarity of the final solution (a. 0.36uM, b. 1.00uM and c. 10pM)
V1 = Initial concentration (???)
V2 = Final concentration (50ul)

Example:
M1:100uM; V1:?; M2:0.36uM; V2:50ul)
M1V1 = M2V2
(100uM)(V1) = (0.36uM)(50ul)
(100uM)(V1) = 18
V1 = (18uM/ul)/(100uM)
V1 = 0.18ul

*** This is applicable if your lab have a 0.1-2.5ul pipette. If you don't have, then dilute the stock primer further to 50uM.***


Best Regards,
Vin

THIRUMURUGAN on Tue Jun 28 06:59:09 2011 said:

Just a minor correction: V1 and V2 refer to VOLUME and not concentration.

The formula can also be noted as: C1V1 = C2V2 where C is concentration and V is volume just make sure to keep units consistent.