NucleoBond
®
Xtra Midi/Maxi -
English
MACHEREY-NAGEL – 01/2008/ Rev. 04
28
Midi
Maxi
13 Precipitation
Add
room-temperature
isopropanol
to precipitate the eluted plasmid DNA. Vor-
tex well and let the mixture sit for
2 minutes
.
Centrifuge at
5,000 x
g
for
15 min
at
room temperature,
preferably
at
15,000 x
g
for
30 min
at
4°C
. Carefully discard the supernatant.
3.5 ml
10.5 ml
14 Wash and dry DNA pellet
Add
room-temperature 70%
ethanol
to the pellet and centrifuge at
5,000 x
g
,
preferably
15,000 x
g
for
5 min
at
room temperature
(20-25°C).
2 ml
5 ml
Carefully remove ethanol completely from the tube with a pipette tip. Allow the
pellet to dry at
room temperature
(20-25°C).
Note: Plasmid DNA might be harder to dissolve when over-dried.
5-10 min
10-15 min
15 Reconstitute DNA
Dissolve the DNA pellet in an appropriate volume of buffer TE or sterile H
2
O. De-
pending on the type of centrifugation tube, dissolve under gentle pipetting up and
down or constant spinning in a sufficient amount of buffer for 10-60 min (3D-
shaker).
Determine plasmid yield by UV spectrophotometry. Confirm plasmid integrity by
agarose gel electrophoresis (see section 4.12).
