I am struggling in doing PCR , it sounds thai I have some problem . the big problem I have been facing is to calculate the the DNA and primers concentration after measuring it by nanodrop.
for example: wHAT IS written in he bottles as follows:
I have DAN
T7 -forward primers:
120.3nm , M w:12.754.3. I have made 100um stock solution by multiple 18.6 *10 += 186 water added
155.6NM M w:5.852.8
I have made 100um stock solution by multiple155.6 *10 and then 1556 water added . I uasually take 10 ul from the stock to add to the 1ML PCR reaction to make 1 um final concentration. however, I did not got any good resuilt yet .
my supervisor told me that I have to make 1fetomolar concentartion from DNA and then use the nanomolar concentration from the both primers ( iam not sure 1 or 10 nanomolar)
how to make this ratio :
is that kind of diluation if so how can i dilute it since i got very unmeasurable number when i use the formula cv=c1v1
what is the right and accurate steps should follow when measuring DNA concentration by nanodro and getting (2.3 ng/ul ) to make the unit nanomolar nMor fetomolar fM
any one can help.