Caroline’s Guide to Quick mini-prepping using the Qiagen Turbo 8 Kit
1. Spin overnight cultures at 4000 rpm for 4 min.
2. Remove media and place tubes in wire rack.
3. Use Finnitips in old 1mL pipette racks and the variable-spaced multichannel pipetman to add 250 uL buffer P1 and resuspend the cultures by pipetting up and down. The Finnitips should be replaced between samples; this must be done by hand.
4. Add 250 uL buffer P2 and mix by gently shaking the entire wire rack.
5. Allow buffer P2 to lyze cells for 5 min. During this time, place the TurboFilter 8 strips in the top plate of the vaccum manifold and place the Qiaprep 8 strip in the plexiglass strip holder directly below the TurboFilter 8 strips. Unused slots should be sealed with blank plexiglass pieces.
6. Add 350 uL buffer N3 to the lyzed cells. Mix by gently shaking the entire wire rack.
*The next several steps are exactly as described in the manual*
7. Transfer the disrupted cells to the TurboFilter 8 strips. Unused wells should be sealed with the strip caps.
8. Apply vaccuum gradually, and allow full vaccuum to pull all liquid into the Qiaprep 8 strips.
9. Switch off vaccuum, and slowly release pressure. Discard TurboFilter 8 strips, and transfer Qiaprep strips to top plate. Replace the plexiglass strip holder with the waste tray. Apply vaccuum slowly to draw cleared lysate through Qiaprep 8 strips.
10. Turn off vaccuum and release pressure.
11. Use electronic multichannel pipette (8 x 1500) with Finnitips (in their normal rack) to wash strips with 1mL buffer PE.
12. Repeat wash step.
13. After washes, apply full vaccuum for 5 min to dry the membrane.
14. Slowly release pressure. Remove top plate, still containing Qiaprep strips, and tap on a stack of Kimwipes until no more drops come out. Blot the nozzles of the Qiaprep strips with a clean Kimwipe.
15. Replace waste tray with blue collection microtube rack containing the 1.2 mL microtubes. Make sure that B1 of the collection tube rack is directly underneath the first Qiaprep strip.
16. Add 100 uL buffer EB to the center of each well using the electronic multichannel pippette (8 x 300 uL). Let stand 2 minutes.
17. Apply maximum vaccuum for 5 minutes.
18. Open the top lid and remove the Qiaprep strips.
19. Remove the blue collection tube rack and centrifuge briefly (2000 rpm, 1 min) to collect liquid in bottom of tubes. Seal with caps.