How do I quantify oligonucleotides by spectrophotometer?
Here is a protocol for the Quantification of Oligonucleotides by Spectrophotometer:
- Add an aliquot of the resuspended oligonucleotide into a volume of PBS so that the total volume is 1000 µl. Typical dilutions are 1:20 or 1:40 where the dilution factor (DF) is 1000/aliquot volume.
- Vortex or pipette up and down repeatedly for 15 seconds.
- Read the absorbance of this dilution at 260 nm (OD260). Use the average of at least 2 reads.
- Calculate concentration using the nmol/OD260 value presented on the Certificate of Analysis, i.e. multiply (nmol/OD260) x (average OD260) x (Dilution Factor) = [C], concentration in µM (micromolarity).