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Primer Notes 1. Primer yield and concentration (approximate )
 

Summary: 52
Primer Notes
1. Primer yield and concentration (approximate ):
1 OD260 of ssDNA 37 g DNA
A260 value X 37 = g oligo
g oligo Mr = mol oligo (X 1000 = nmol oligo)
Average nucleotide 325 Daltons
Example: 10 OD260 units of a 20-mer in 1 ml 57 nmol/ml 57 M 57 pmol/l
Primer yield and concentration (exact):
g/OD260 = MW (ammonium salt)/extinction coefficient
MW (ammonium salt) = MW (oligo) + [(# of bases -1) X 16.06]
MW (oligo) = [(#A X 313.2) + (#C X 289.2) + (#G X 329.2) + (#T X 304.2) + (#U X 290.2)
+ (#I X 252) + (#N X 309)] - 62
Extinction coefficient = (#A X 15.3) + (#C X 7.4) + (#G X 11.8) + (#T X 9.3) + (#U X 10.1) +
(#I X 12.25) + (#N X 10.95)
2. Primer storage:
It is best to store primers at a high concentration at -20C. Typically, this will mean 50X -100X
(or more) concentrated than used in sequencing or PCR reactions. Thus, it will be necessary to
dilute the primer stock solution before use. Keep the dilute primer at -20C until it is exhausted.
3. Reconstitution:

  

Source: Aris, John P. - Department of Anatomy and Cell Biology, University of Florida

 

Collections: Biology and Medicine