DNase Protocol

Overview
Protocol for DNase treatment of total RNA. This uses a DNase inactivating reagent and thus avoids the DNase heat inactivation step that makes me nervous.

Materials

 * The DNAfree kit contains the stuff you need: Applied Biosystems
 * THAWED 10X DNase Buffer

Procedure

 * 1) In order to DNase 20ug of RNA in a 100ul reaction, mix the following:
 * 10&mu;L 10X DNase buffer
 * 78&mu;L dH20
 * 2&mu;L DNase
 * 10&mu;L RNA(1ug/&mu;L)


 * 1) Incubate at 37&deg; C for 20-30 min.
 * 2) Add 0.1 volume resuspended DNase Inactivation Reagent.
 * 3) Incubate 2 min at RT mixing every 30s.
 * 4) Centrifuge at 10,000xg for 1.5 min.
 * 5) Transfer solution to a new tube avoiding pelleted Inactivation Reagent.

Contact

 * Who has experience with this protocol?

or instead, discuss this protocol.