CTAB Procedure

In the hood:
96 well dish with bacteria
titertech
microtubes
glass pipette

Remove colonies from each well using the titertech and place them into the cover
Pipette up and down to thoroughly mix the colonies
Aliquot 300 %26micro;l of the culture into a microtube; save about 4 or 5 tubes

In the lab:
Centrifuge the culture for about 2 minutes or until a pellet has formed
Remove the supernatant
Resuspend the pellet in 576 %26micro;l TE, 15 %26micro;l of 20% SDS and 3 %26micro;l of 20 mg/ml proteinase K
Incubate for 1 hour at 37 ℃
Add 166 %26micro;l of 3M NaCl and mix thoroughly
Add 80 %26micro;l of 10% CTAB in 0.7 M NaCl and thoroughly mix
Incubate for 10 minutes at 65 ℃

In the hood in the back:
Add an approximately equal volume of chloroform (700 %26micro;l)

In the lab:
Centrifuge for 5 minutes at room temperature

In the hood in the back:
Remove white interface (should be able to be done by using pipetter)
Transfer supernatant to another microtube
Discard remaining solution in chloroform waste receptacle
Add an equal amount of phenol/chloroform

In the lab:
Centrifuge for 5 minutes

In the hood in the back:
Transfer supernatant to a new microtube
Discard remaining solution in proper waste receptacle

In the lab:
Add 0.6 vol of isopropanol and gently rock back and forth until white precipitant forms
Centrifuge for 30 minutes
Remove supernatant and wash pellet with 70% ethanol
Centrifuge the tube for 5 minutes
Remove supernatant
Redissolve the pellet in 100 %26micro;l TE/10