Sean Lauber:DNase Treatment Using DNA-Free
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Procedure
Using Ambion DNA-free (CAT# 1906)
- 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.
- Add 2.5 μL of DNAse buffer I
- Add 1 μL of DNAse I, tap tube to mix gently
- Incubate at 37*C for 30 min
- Add 5 μL DNAse Inactivation Reagent, tap tube to mix
- Incubate at RT for 2 min, tapping occasionally to ensure the solution is mixed
- Spin 10K RPM/RT/1 min
- 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