How do I figure out the concentration of my oligo?
A. Your oligo supplier includes an oligo specification sheet when they ship your oligo. The information on the spec sheet includes: oligo name, oligo sequence, # bases, molecular weight (MW), nmols/OD260, g/OD260, GC content, and Tm (50mM NaCl). Also the amount of oligo synthesized is reported in terms of OD260, nanomoles and mg. Using the the nanomoles of oligo synthesized and the desired concentration of the primer, you can calculate the amount of TE to add to make a 100 uM stock solution. NOTE: Always resuspend oligo stocks in TE. (mols of primer/ desired concentration of primer) = volume of TE to resuspend primer (87.16 nmols / 100 uM) = 871.6 ul (87.16 x 10-9 mols / 100 x 10-6 mols/L) = 0.8716 x 10-3 L = 871.6 ul of TE Now that you have your 100 M oligo stock solution, you need to dilute a portion of it to the correct primer concentration for sequencing your plasmid, 1.6 M. uM is equivalent to pmol/ L. 1 pmol/ ul = 1 x 10-12mols /1 x 10-6 ul = 1 x 10-6 mols/L= 1 uM. To make 100 ul
