electroelution protocol

2013-12-01

azim58 - electroelution protocol

Electroelution protocol

see
Electroelution with Elutrap

referred to protocol here
http://humgen.wustl.edu/hdk_lab_manual/rdm/rdm9.html

Note: minimize exposure to UV (use prep (long form) of UV to cut out the
DNA
Note: expected yield is 50-75%

Cut a piece of dialysis tubing approximately 3 cm longer than the gel

slice and clip one end. Gently push gel slice into open end and down to
the clip. Add some buffer

Place gel slices parallel to the electrodes and fill the electrophoresis

box with buffer (again, same as original gel) until all tubing is
submerged.

Electroelute at 80-100 volts for 2-5 hours, longer for large fragments.

The movement of the DNA can be monitored with a hand held UV source
(long-wavelength).

DNA may stick to dialysis tubing after current flow. Remove the gel

piece from the dialysis tubing. Reverse the electrodes and run for 15 s
to 2 min to dislodge the DNA from the membrane surface.