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-   -   Maybe stupid question..(primers) (http://www.molecularstation.com/forum/basic-lab-protocols-techniques/48094-maybe-stupid-question-primers.html)

ineska 01-10-2009 10:32 PM

Maybe stupid question..(primers)
 
hi there..
it's a little bit stupid question but I have to make sure..
about primer concentration:
I have 30nM of lyophilized primer, I add 300 ul water or TE buffer and get 100uM/L stock solution. If I add 600 ul water I get 200uM/L stock solution???
So to prepare 10uM/L work solution I have to take 20ul primer stock solution and add 80 ul water?? Am I right? Question is cause I did the dilution one more time after did it the first one...
thank You in advance!!!

butters 02-06-2009 04:04 AM

Re: Maybe stupid question..(primers)
 
30nM + 300ul water = 300ul of 100uM stock solution.

*if 30nM + 600ul water = 600ul of 50uM stock solution
**remember what ever amount of water you added you only have 30nM of primer

To prepare 10uM of primer you need
1 in 10 dilution for 100uM (10ul + 90ul of water)
or
1 in 5 dilution for 50uM (20ul + 80ul of water)

if you add 600ul then u would need from stock 20ul + 80ul of water.


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