96-well DNA Extraction Protocol from 25mm 0.2µm filters
Prepare lysis buffer. 
Prepare Lysozyme & RNase Aliquot. 
Prepare ProK Aliquot. 
Thaw filters on ice. 
Do quick spin down. 
Transfer each filter to screw top, O-ringed eppendorf tube, also on ice. 
Add 250 µl lysis buffer with RNase and lysozyme to each tube. 
Incubate 37°C for 30 min., rotating end over end at angle, for optimal mixing with minimal frothing. 
Add 18.75 µl of Proteinase K solution (10 mg/ml made up in lysis buffer) to a final conc. of 0.65 mg/ml. 
Add 29.9 µl 10% SDS to a final conc. of 1%. 
Incubate at 55°C for 2 hours, rotating end over end at angle. 
Towards end of this time, turn on heat block or hyb oven to 70°C. 
Put elution liquid (Buffer AE or water) into 70°C to preheat. 
Add 300 µl Buffer AL (=Buffer AL/E without the ethanol added). 
Mix thoroughly by vortexing and spin down quickly.  
Incubate 70°C for 10 mins.
Add 300 µl 96-100% EtOH. 
Mix by vortexing vigorously and spin down quickly. 
Check pH of lysate, must be <7 to get max. binding efficiency to column. 
Pipet onto 96 well spin columns, making sure not to wet the rims to avoid cross contamination. 
Place the 96 well column plate onto the S-block for flow through collection. 
Seal plate with Airpore tape sheet Spin 5788 xg for 10 min. at 40­°C. 
Discard flow through. 
Place in new collection tray.  
Add additional lysate if needed and repeat spin, etc.
Add 500 µl Buffer AW1, reseal plate Spin 5788 xg for 5 min. at 40°C. 
Add 500 µl Buffer AW2, reseal plate Spin 5788 xg for 5 min. at 40°C. 
To dry columns, either reseal plate with new sheet and spin 5600 xg for 15 min. at 40°C atop a new collection tray or incubate in hyb oven at 70°C for 15 min.
Transfer column plate to top of rack of "elution microtubes RS". 
Add 200 µl pre-heated 70°C Buffer AE or water, reseal plate Incubate 1 min. at RT.  
Spin 5788 xg 2 min. to elute. 
Repeat with second 200 µl (will increase total yield up to 25%) or if you wanted to keep the column small, you could use the first 200 µl elution, heat it back to 70°C, and pass it through the column again (will increase total yield approx. 15%). 
Can freeze elutants and break here overnight, or for a while, before proceeding with Final DNA clean up step, particularly if DNA is in Buffer AE which is TE, so will keep the DNA relatively stable. 
Transfer the eluted DNA to the 96 well PCR purification plate (no more than 300 µl at a time). 
Apply vacuum at 20 inches Hg until dry (~20-30 min.)
Rinse DNA with 100 µl Ambion water, apply vacuum 5-10 min. until dry. 
Rinse a second time with water if you want to make sure all Buffer TE removed. 
Add 20 µl dilute TE. 
Pipette up and down 20 times and transfer to a clean 96 well plate
