Sean Lauber:DNase Treatment Using DNA-Free

Procedure

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