I have been preparing my primer solutions this way and need some help
Here is an example of 1 primer tube and how I've made a working solution
Primer M.W= 6,298.1
I first suspend the primers in 100ul of PCR grade H2O= 250ug/100ul=2.5ug/ul
The PCR optimization guide told me to add 1ul of (0.25ug/ul) working solution to my reaction.
To get my 0.25ug/uL primer working solution of 200ul total volume I take 20ul of stock and 180ul of PCR grade H20 v1(2.5)=200(0.25)
1.) I want to know what the uM concentration of this solution is, because the gene sequencing lab is requesing primer solutions of 2uM.
2.) Also my PCR optimization guide says primers should be between 0.1uM and 0.5uM final concentration in my PCR solutions. In my case 50uL total PCR reaction solution. What would be the final uM concentration for this primer?
Many many thanks