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| PCR - Polymerase Chain Reaction Forum 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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#1
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| Hi everyone I have a 100然 Primer Stock which I'm going to dilute 1:10 giving me a 10然 working primer soln. I need 20pmol of primer for a PCR reaction that has a final volume of 100痞. Can't figure out how much primer to add. Can anyone help? Thanks |
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#2
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| 100然 corresponds to 100pmol/無, if you dilute it 1/10 then you can simply use 2無 per PCR. |
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#3
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| As Multiplexion has written, 100uM stock that you have got means 100pmol/uL and when you dilute it to 1:10, then your working concentration becomes 10pmol/uL. If you add 2uL it means you have added 20pmol/uL of primer to a reaction of 100uL. |
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#4
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| Hi, its very simple, here is the calculation you have 100uM stock = 100pmols/uL, you did 1:10 dilution so stock concentration became 10uM = 10pmols/uL; Intial concentration (C1) = 10uM = 10pmols/uL how much to take (V1) = ?? Final concentration (C2) = 20pmols/100uL = 0.2pmols/uL Final Volume (V2) = 100uL C1*V1 = C2*V2 10*V1 = 0.2*100 V1 = (0.2*100) / 10 V1 = 2 uL Hope you understood. |
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