8
Fig. 5. Aurum plasmid mini protocol overview: vacuum format
Growth and Isolation
1.
Grow 1–5 ml bacterial culture overnight or
16 hr.
2. Measure
A
600
(if higher yield required).
3.
Transfer an appropriate volume of culture to
a capped 2 ml tube. Centrifuge and
decant supernatant.
4.
Add 250 µl resuspension solution; vortex.
5.
Add 250 µl lysis solution; invert 6–8x.
6.
Add 350 µl neutralization solution; invert 6–8x.
7.
Centrifuge 5 min to pellet cell debris.
Purification on Aurum or
Comparable Manifold
(See exploded view for proper setup of
manifold.)
8.
Transfer cleared lysate (supernatant) to mini
spin/vac column.
9.
Apply vacuum at -20 to -23" Hg to bind
plasmid DNA. Turn vacuum off.
10. Add 750 µl wash solution and reapply
vacuum until all liquid has passed through
column.
11. Transfer mini spin/vac column to a 2 ml wash
tube. Spin
1 min
to remove residual wash.
Collection of Purified Samples
12. Transfer mini spin/vac column to a clean
1.5–2.0 ml capped tube.
13. Add 50 µl elution solution. Let stand 1 min
and then centrifuge 1 min to elute.
14. Purified DNA is ready to use or can be
stored at 4°C.
Bulletin 2663 US/EG Rev B 01-568 0901
4110116 Rev A
Aurum Plasmid Mini Kit: Cat. #732-6400
For more information, call Technical Services
at 1-800-424-6723.
350 µl
neutralization
solution
250 µl
lysis solution
250 µl
resuspension
solution
Aurum
™
Plasmid
Mini Kit:
Vacuum Format
Protocol Overview
For complete protocol, consult instruction manual.
Invert 6–8x
Vortex
Invert 6–8x
50 µl
elution
solution
Transfer
cleared lysate
750 µl
wash solution
™
