Hello colleagues, I need final working concentration of 10uM for each primer (from 100uM stock) but the company has set specification on the amount of primer from 0.025 µmole to 1 µmole. what does this mean? and how will this affect the final stock concentration?
Hello,
As I know that, usually the stock concentration of primer is 50uM, some rare case they make on 100uM. For my case I usually dilute 5 times and then use for PCR.
I think you can just dilute 10 times from your stock and use it for PCR.
Thank you Nam! my problem is on the synthesis scale whether I need to order 0.025 umoles or 1 umoles?
Dear Gaspary
can you send the primer data sheet ? I am wondering if you are confusing concentration with yield ?
I should be able to help you if I can see the company data sheet
EXAMPLE:
If you have 12000 pmol is ugual to 12 nmol.
100 uM means 100nmol/1 ml
x= 12/100
x= 0,12 ml 120 ul (volume of water I need adding to obtain 100 uM)
If you have 100uM stock, then you need to dilute it 10 times.
If your primers is solid (lyophilized), then you need to do some calculations: see Giorgia Pertile answer.
The company is giving you options for the amount of primer molecules in moles. This is an amount, and not a concentration. Your primers will most likely come as a dry powder and then you add water to bring it up to what ever final volume is needed to create your desired final concentration. Ask your lab colleagues what amount they usually order, they probably have a preference based on price and anticipated use.
Dear Laurence Stuart, find the attached primer data sheet last column is amount (synthesis scale) give as micro moles
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