Bio-Rad Aurum™ Plasmid Mini Purification Module - Manual (Page 8)

Section 6
Protocol
Vacuum Format
This procedure requires the Bio-Rad Aurum vacuum manifold and column
adaptor plate (Cat. #732-6470), or any vacuum manifold with luer fittings. For
proper vacuum setup conditions, please read Instrument Setup and Use
for the Column Adaptor Plate in the vacuum manifold instruction manual,
and see Fig. 4. All centrifugation steps are performed at maximum speed
( 12,000 x g) under ambient conditions, with any commercially available
microcentrifuge which can accommodate 1.5 ml and 2.0 ml tubes. Please read
the section "Guidelines for Using the Aurum Plasmid Mini Kit" before
proceeding.
1. Transfer up to 12 OD•ml of plasmid-containing bacterial host to a 1.5–2.0
ml capped microcentrifuge tube (not provided). Pellet the cells by
centrifugation for 1 min. Remove all supernatant by decanting or pipetting.
2. Add 250 µl of resuspension solution and vortex or pipet up and down until
the cell pellet is completely resuspended.
3. Add 250 µl of lysis solution and mix by inverting the capped tube briskly
6–8 times. DO NOT VORTEX OR SHAKE. The solution should become
viscous and slightly clear.
5
Fig. 2. Vacuum regulator with clear representation of the gauge