Protocol #01
DNA isolation
Using UltraClean Kit form QIAGEN for isolating DNA from swabs
Prep:
 01 If precipitated, heat solution SL at 55 °C until precipitates dissolve.
 02 Vortex the collection tube with the swab for several seconds, follow by quick spin.
 03 Transfer 300 µL of the supernatant from the collection tube to the PowerBead tube.
 04 Add 50 µL of Solution SL to the PowerBead tube. Vortex the mixture.
 05 Incubate the PowerBead tube at 70 °C for 5 minutes. Quick spin afterwards.
 06 Secure PowerBead tubes horizontally on the plate vortex using a red box with tape.
Vortex at maximum speed for 10 minutes.
 07 Centrifuge the tubes at 10 000 g for 1 minute.
 08 Transfer the supernatant into a new 2 mL tube.
 09 Add 100 µL of Solution IRS and vortex for 5 s. Incubate at 4 °C for 5 minutes.
 10 Centrifuge the tubes at 10 000 g for 1 minute.
 11 Transfer the supernatant into a new 2 mL tube. Avoid the white pellet.
 12 Add 900 µL of Solution SB and vortex for 5 s. Shake the Solution SB before use.
 13 Transfer 700 µL of the mixture from step 12 into an MB Spin Column.
Centrifuge at 10 000 g for 1 minute. Mark both the column and the collection tube.
 14 Discard the flow-through. Transfer another 700 µL of the mixture from step 12 into an
MB Spin Column. Centrifuge at 10 000 g for 1 minute.
 15 Discard the flow-through. Transfer the remaining mixture from step 12 into an MB
Spin Column and centrifuge again at 10 000 g for 1 minute. Discard the flow-through.
 16 Add 300 µL of Solution CB into the MB Spin Column. Shake the Solution SB before use.
Centrifuge at 10 000 g for 1 minute.
 17 Discard the flow-through. Place the MB Spin Column in a new 2 ml collection tube.
Centrifuge at 10 000 g for 1 minute.
 18 Discard the flow-through. Place the MB Spin Column in a new 2 ml collection tube.
Be careful not to splash any of the remaining liquid on the MB Spin Column.
 19 Add 50 µL of Solution EB to the centre of the white filter membrane.
 20 Incubate at RT for 5 minutes.
 21 Centrifuge at 10 000 g for 1 minute.
 22 Discard the MB Spin Column and keep the flow-through.