Isolation of DNA from phage lysate
Put phage lysate in plastic centrifuge bottle and add RNase ( for 100 ml add 10 µl , for 150 ml add 15 µl , etc . )
Incubate 30 min . at room temperature .
Add NaCl to a final concentration of 1M ( for 100 ml add 6.5 g NaCl ) .
Incubate for 1 hour on ice Centrifuge for 10 min . at 11,000xg ( 8,300 rpm on GSA rotor ) .
Transfer supernatant to a new bottle .
Add PEG 8000 at 100g / l ( for 100 ml add 10 g PEG ) and shake to mix .
Incubate 1 hour on ice Centrifuge for 10 min . at 10,000xg ( 7,835 rpm on GSA rotor ) .
Carefully pour out the supernatant to discard .
Set the bottle upside down on paper towels / Kimwipes to drain remaining liquid .
Rinse the inside of the bottle twice with SM buffer ( total volume of 1 - 2 ml ) .
Collect in 2 ml microcentrifuge tubes ( 1 ml per bottle ) .
Shake resin to resuspend and heat TE to 80°C .
Add 1 ml resin to each 2 ml tube .
Mix by inversion .
Attach column to 3 ml syringe and push sample through 1 ml at a time .
Push 2 ml of 80 % isopropanol through each column 1 ml at a time .
Place column in original 2 ml tube .
Centrifuge 2 min at 10,000xg to remove excess isopropanol .
Place column in new 1.5 ml microcentrifuge tube .
Add 100 µl warm TE to elute .
Briefly vortex Centrifuge for 30 sec . at 10,000xg .
Check concentration of isolated DNA using NanoDrop .
Use Quant IT DNA quantification to validate NanoDrop readings
