Centrifuge at high speed in a microcentrifuge for 5 minutes. Vortex the sample for about 2 seconds, wait for 5 minutes, vortex for another 2 seconds, and separate the layers with a 5 minute spin in a microcentrifuge. Collect, then extract the upper aqueous phase with ether 3-4 times to remove phenol. Spin for 5 minutes and dry the pellet under vacuum. Allow reaction to proceed for 15 minutes at room temperature.
Timing is critical.
Allow to react at room temperature for 15 minutes. QS to 500 ml with H 2 O and autoclave. Molecular Biology Protocols .