Sean Lauber:DNase Treatment Using DNA-Free

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Using Ambion DNA-free (CAT# 1906)

  1. Bring your RNA solution up to a final volume of 21.5 μL. Be sure that your RNA solution contains less than 10 μg of total RNA.
  2. Add 2.5 μL of DNAse buffer I
  3. Add 1 μL of DNAse I, tap tube to mix gently
  4. Incubate at 37*C for 30 min
  5. Add 5 μL DNAse Inactivation Reagent, tap tube to mix
  6. Incubate at RT for 2 min, tapping occasionally to ensure the solution is mixed
  7. Spin 10K RPM/RT/1 min
  8. Pipette 15 μL of the supernatant into a new tube. Take more if you dare, but you don't want to transfer over the Inactivation Reagent