Macherey-Nagel NucleoSpin 740727.10 User Manual page 15

4 Bind DNA
Place a NucleoSpin
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.
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.
6 Elute DNA
Place the NucleoSpin
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.
NucleoSpin Plasmid EasyPure
®
Plasmid EasyPure Column into a
®
Plasmid EasyPure Column in
®
MACHEREY-NAGEL – 08/ 2 013, Rev. 01
Load
supernatant
1,000–
2,000 x g,
30 s
+ 450 μL AQ
> 12,000 x g,
1 min
+ 50 μL AE
RT, 1 min
> 12,000 x g,
1 min
15