Troubleshooting guide

QIAprep Spin
QIAprep Miniprep Handbook 12/2006 25
During centrifugation, prepare the vacuum manifold and QIAprep spin columns:
QIAvac 24 Plus (see pages 16 and 18–19):
■ Ensure that the vacuum source is connected to the upper threaded hole of the
QIAvac 24 Plus and the lower threaded hole is tightly sealed using the screw
cap.
■ If using the QIAvac Connecting System, connect the system to the manifold
and vacuum soured as described in the
QIAvac 24 Plus Handbook .
■ Insert up to 24 spin columns into the luer slots of the QIAvac 24 Plus. Close
unused luer slots with luer plugs.
QIAvac 6S manifold:
(Note: The following procedure applies to the manifold with a hinged lid and
spring lock. See pages 16 and 18–20.)
■ Open QIAvac 6S lid. Place QIAvac luer adapter(s), or blanks (provided with
QIAvac 6S) to seal unused slots, into the slots of the QIAvac top plate. Close
the QIAvac 6S lid. Place the waste tray inside the QIAvac base, and place
the top plate squarely over the base. Attach the QIAvac 6S to a vacuum
source.
■ Insert each QIAprep spin column into a luer connector on the luer adapter(s)
in the vacuum manifold. Seal unused luer connectors with plugs provided with
the QIAvac Luer Adapter Set.
Other vacuum manifolds: Follow the supplier’s instructions. Insert each QIAprep
column into a luer connector.
5. Apply the supernatant from step 4 to the QIAprep spin column by decanting or
pipetting.
6. Switch on vacuum source to draw the solution through the QIAprep spin columns,
and then switch off vacuum source.
7. Recommended: Wash the QIAprep spin column by adding 0.5 ml Buffer PB. Switch
on vacuum source. After the solution has moved through the column, switch off
vacuum source.
This step is necessary to remove trace nuclease activity when using
endA
+
strains
such as the JM series, HB101 and its derivatives, or any wild-type strain, which
have high levels of nuclease activity or high carbohydrate content. Host strains
such as XL-1 Blue and DH5α do not require this additional wash step.
8. Wash the QIAprep spin column by adding 0.75 ml Buffer PE. Switch on vacuum
source to draw the wash solution through the column, and then switch off vacuum
source.