DNA extraction from plants
Grind frozen plant material (100mg) in liquid Nitrogen.
In a 2 ml tube, add 800 uL of 1,5x CTAB and 1 ul of Beta-mercaptoethanol to the ground leaf material.
Incubate 1 hour at 60-65 degrees (C).
Cool at Room Temp.
Add 1 volume of Chloroform/Isoamylalkohol (24:1) mixture.
Mix on overhead shaker for 10 minutes.
Centrifuge at 3000 rmp for 25 minutes.
Transfer supernatant to a new tube.
Use wide pipette tips (cut point of tips if needed).
Add 10 uL RNAse (10 mg/ml) incubate at 37 degrees (C) for 30 minutes.
Add one volume 100% Isoponanol (pre-chilled at -20).
Allow sample to precepitate for up to 20 minutes at -20.
Centrifuge at full speed for 10 minutes.
Remove supernatant.
Add 1 ml 70% Ethanol.
Centrifuge at full speed for 10 minutes.
Remove Supernatant.
Repeat once.
Dry pellet in air or approx 20 minutes in vacuum centrifuge.
Dissolve pallet in 1x TE (10 mM Tris, pH8, 1 mM EDTA)
