DNEasy DNA Extraction - Vibrio Gram Negative Broth Bacteria
Put Buffer ATL and AL at 56 C for 5 minutes before using.
Transfer 250ul of broth culture to 2ml deep well (autoclaved). 
Harvest cells (maximum 2 x 109 cells) by centrifuging plate for 15 min at 4063 x g. 
Discard supernatant.
Resuspend pellet in 200 µl Buffer ATL by pipetting up and down 15 times.
Seal with foil and place in oven at 56 C for one hour.
Place extra plate on seal so foil doesn’t come off.
Pipette up and down 15 times and spin down.
(May place in -80 C and continue procedure later at this point.)
If placed in -80 C, let thaw and spin down @ 1000g for 1 min. 
Add 205 uL Buffer AL and 205 uL molecular grade EtOH.
Pipette up and down 15 times and spin down.
Place DNeasy 96 plates on top of 2ml deep well plate .
Mark the DNeasy 96 plates for later sample identification.
Carefully transfer the lysate (approximately 600ul) of each sample from step 7 to each well of the DNeasy 96 plates.
Do not transfer more than 900 µl per well.
Seal each DNeasy 96 plate with a porous film.
Centrifuge for 15 min at 4063g.
If lysate remains in any of the wells, centrifuge for a further 10 min.
Add 500 µl Buffer AW1 to each sample.
Seal each DNeasy 96 plate with a new AirPore Tape Sheet (provided).
Centrifuge for 5 min at 4063g.
Remove the tape.
Carefully add 500 µl Buffer AW2 to each sample.
Centrifuge for 15 min at 4063g.
Do not seal the plate with AirPore Tape.
The heat generated during centrifugation ensures evaporation of residual ethanol in the sample (from Buffer AW2) that might otherwise inhibit downstream reactions.
Place in new collection rack (2ml deep well plate) and spin again 15 minutes at 4063g.
Place each DNeasy 96 plate in the correct orientation on a new rack of VWR 500ul plate.
To elute the DNA, add 200 µl Buffer TE to each sample, seal and incubate for 1 minute at room temp.
Centrifuge for 4 min at 4063g.
200 µl Buffer TE is sufficient to elute up to 75% of the DNA from each well of the DNeasy 96 plate.
Recommended: For maximum DNA yield, repeat step 16 with another 200 µl Buffer TE.
Aliquot 20ul of extracted DNA to green 96 microplate.
Seal DNeasy 96 plate and 96 microplate with non-sterile foil.
Place DNeasy 96 plate in -80 C and 96 microplate in -20 C.
