DNAse 1 Protocol

From OpenWetWare
Jump to navigationJump to search

DNA-free Protocol

Routine Protocol (20ug RNA in a 100ul rxn)

1) Mixup the following:

10X DNAse Buffer 10ul dH20 70ul DNAse 2ul RNA(1ug/ul) 20ul

2) Incubate at 37C for 30 minutes 3) Add 0.1 volume DNAse Inactivation Reagent (i.e. 10ul) 4) Incubate at room temperature for 2 mins mixing every 20-30 seconds. 5) Centrifuge at 10,000xg for 1.5 minutes at room temperature. 6) Pull off liquid without disturbing the pelleted Inactivation Reagent and transfer to a new tube.