Quantitation of DNA

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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