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Modif ied f rom IDT Technical Bulletins Everything you ever wanted to know concerning Oligonucleotides but were afraid to ask Calculating Oligonucleotide Quantity Specifications Sheet Calculations and Conversion Factors The specific quantity of an oligonucleotide is obtained by the following set of formulas and conversion factors. The absorbance at 260 nanometers, OD260 , of each oligonucleotide is read at two different dilutions. The value reported on the specification sheet is the average of these two measurements. The number of milligrams and nanomoles of the oligonucleotide is calculated from the OD260 reading using the following formulas: For single stranded DNA: 1 OD260 = 33 ug = 0.033mg milligrams = OD260 x 0.033 6 nanomoles = [milligrams ÷ molecular weight] x 10 The molecular weight of each oligonucleotide is presented on that oligo’s specification sheet. Calculation of Molecular Weight for Synthetic Oligonucleotides The molecular weight of an oligonucleotide is calculated based on the following formula: Molecular Weight (gm/mole) = (251 x nA) + (245 x nT) + (267 x nG) + (230 x nC) + (61 x (n-1)) + (54 x n)+(17 x (n - 1)) + 2 Where: 1) nA = number of dA bases 2) nT = number of T bases 3) nG = number of dG bases 4) nC = number of dC bases 5) n = total number of bases in the sequence 6) (61 x (n - 1)) accounts for the molecular weight of the phosphate groups. 7) (54 x n) accounts for the hydration of the DNA. There are approximately three water molecules per nucleotide. 8) (17 x (n - 1)) accounts for the ammonium cations associated with the phosphate groups. Sample Calculation: Example sequence: ATG TAA TGT TTG GTC CGC nA=3, nT=7, nG=5, nC=3, n= 18 Using th