Quantitation of DNA

HOW TO ... quantitate DNA, Rutgers, December 2004

1. Equipment needed
   • UV spectrophotometer
   • 100 ul UV cell
   • vortex
   • spin
2. Reagents needed
   • H2O DNA grade
3. Other consumables
   • 2.5 ml tubes, sterile
   • gloves
   • autoclaved/sterile tips
4. Procedure
   • dilute samples 100 times (2 ul product + 198 ul H2O)
   • read OD at 260 and 280 nm, reading time should be > 6 sec
   • use the average of the two readings
   • OD260 of 1 corresponds to a concentration of 50 ng DNA/ul or to 40 ng RNA/ul
   • OD260/OD280 should be higher than 1.6, a lower value indicates the presence of proteins