| | Re: Real Time PCR dilution
I am confused about your working stock conc that primer was at 300nmol and probe at 100nmol, most working stock is from 5Ám/Ál to 40Ám/ul, did you mean the final primer/probe reaction conc( add 0.5ul primer or probe to 25ul final reaction), if so, you should make primer working conc as 15Ám/ul and probe as 5Ám/ul,
add 100Ál water to your original stock conc primer (10nmol) to gain 100Ámol Further Stock Conc (FSC) primer, then dilute 15Ál FSC primer into 85Ál water to gain 15Ám/ul.
dilute your stock conc probe as 20 times to gain 5Ám/ul working conc probe.
Last edited by namishu95; 12-22-2010 at 03:59 PM.