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 )
