GE PhastSystem, Руководство пользователя

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Trouble Shooting Guide
This guide lists the symptoms, in illustrated form
(when possible), of some problems you might
encounter with separation and development
techniques. Probable causes are given along with
solutions to correct or prevent the problems. The
symptoms, drawn in gel format, are amplified so
you can easily identify them, and refer to them.
Some symptoms, such as missing bands, are
impossible to illustrate, so the symptoms is
described in the gel outline.
This guide is divided into two major parts:
separation problems and development problems.
Each part is subdivided into problem common to
both PhastGel IEF, homogenous and gradient
media and problems specific to each of these
media (when applicable). The development section
includes trouble shooting guidelines for both
coomassie and silver staining. Note: This guide is
specifically concerned with the problems that
might occur with the techniques presented in the
technique files, e.g. coomassie staining with
Phastgel Blue R. These guidelines may or may not
pertain to other methods using different reagents.
Separation
Symptom Solution(s)
1. Remember to remove the film
before starting a run.
2. Check the electrodes to make sure
they are even. Gently bend them
down a little. Always check the
electrode contact with gel or
buffer strips.
3. Check that the sample applica-
tors, gel cover and buffer strip
holder are positioned correctly.
1. Remove excess water after posi-
tioning gels onto the separation
bed. Use only about 60-75 µl un-
der each gel.
Increase the temperature of the
bed if the humidity in your lab is
too high.
2. Make sure the gel is positioned
correctly within the red vertical
outline on the bed.
Take care you when you insert the
sample applicator, especially when
using the buffer strip holder.
Probable Cause
1. The plastic film on the gel surface
was not removed.
2. No electrode contact with the gel.
3. The sample was not applied to the
gel.
1. Excess water on the bed sur-
rounds and floods the gel.
2. Samples were applied too close to
the gel´ s edge.
Sample list on the plastic when you
put the sample applicator in the slot.
Occasional
empty lanes
No bands
on the gel
Trouble Shooting Guide
Disturbances at one
or both of the edges.