15
MACHEREY-NAGEL – 08/ 2013, Rev. 01
4 Bind DNA
Place a NucleoSpin
®
Plasmid EasyPure Column into a
Collection Tube (2 mL) and decant the supernatant from
step 3 onto the column.
Centrifuge for 30 s at 1,000–2,000 x
g
.
Discard flow-through and place the spin column back into
the collection tube.
Load
supernatant
1,000–
2,000 x
g
,
30 s
5 Wash and dry silica membrane
Add
450 μL Buffer AQ
(supplemented with ethanol, see
section 3).
Centrifuge for 1 min at full speed (> 12,000 x g).
Very carefully discard the collection tube and the flow-
through and make sure the spin cup outlet does not
touch the wash buffer surface. Otherwise repeat the
centrifugation step.
Note: To reduce ethanol carry-over to a minimum for better
performance in downstream applications, incubate spin cup
for 10–15 min at 37 °C to dry silica membrane completely.
+ 450
μL AQ
> 12,000 x
g
,
1 min
6 Elute DNA
Place the NucleoSpin
®
Plasmid EasyPure Column in
a 1.5 mL microcentrifuge tube (not provided) and add
50 μL Buffer AE
.
Incubate for 1 min at room temperature (18–25 °C).
Centrifuge for 1 min at full speed (> 12,000 x g).
Note
: For more efficient elution procedures and alternative
elution buffer (e.g., TE buffer or water) see section 2.4.
+ 50
μL
AE
RT, 1 min
> 12,000 x
g
,
1 min
NucleoSpin
®
Plasmid EasyPure
