19
A protocol for In-Gel Westerns is provided in the Odyssey
®
Appli-
cation Protocols manual. Coomassie-stained gels can also be
scanned since Coomassie Blue dye can be seen clearly in the 700 nm
channel, and faintly in the 800 nm channel (see the Western Blot
Analysis protocol for details). As well, nucleic acids stained with
Syto
®
60 and separated in a gel can be imaged in the 700 nm channel
(see the Syto 60 Staining of Nucleic Acids in Gels protocol for more
information). To scan a gel, follow these procedures:
1) Thoroughly rinse the gel with destaining solution or water to
remove dye particulates.
2) When placing the gel on the scanning surface, take care not to
trap air bubbles underneath. Cover the gel with plastic wrap to
prevent drying, if desired.
3) Scan the gel in the 700 nm channel.
4) Adjust the focus offset for the gel thickness. The correct focus
offset is 1/2 the thickness of the gel; for a 1 mm gel, set the focus
offset to 0.5 mm. The maximum offset is 4 mm in the most current
edition of the Odyssey Infrared Imaging system, allowing gels of
up to 8 mm to be scanned.
Note:
Early versions of the Odyssey Imager were limited to a 2.0 mm focus offset, but
can be upgraded by installing the Odyssey Server Software version 2.0.0 or above.
5) After removing the gel, clean the glass surface to remove any
residual dye by following the instructions in the section
Before
You Begin...
above.
Using Gels
