Chapter 12: troubleshooting, Troubleshooting – Bio-Rad Mini-PROTEAN® TGX™ Precast Gels for 2-D Electrophoresis User Manual

Page 35

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29

Troubleshooting

12

Problem

Cause

Solution

Current is zero or less than
expected, and samples do not
migrate into gel

Tape at bottom of cassette not removed Remove tape

Insufficient buffer in inner buffer
chamber

Fill buffer chamber with running buffer

Insufficient buffer in outer buffer
chamber

Fill inner and outer chambers to ensure
wells of the gels are completely covered

Incorrect cassette orientation

Ensure shorter plate is facing gasket

Electrical disconnection

Check electrodes and connections

Gels run faster than expected

Running buffer too concentrated or
incorrect

Check buffer composition

Gel temperature too high

Do not exceed recommended running
conditions

Gels run more slowly than expected

Companion running module left in tank
when running only 1–2 gels

Remove companion running module

Buffer leaking from inner chamber

Incomplete gasket seal

Wet gasket with running buffer before use

Improper assembly of the gel into the
electrode/companion module

Top edge of short plate should fit under
notch at top of gasket

Top of short plate should touch green
gasket

Bands “smile” across gel: band
pattern curves upward at both
sides of gel

Excessive heating of gel

Check buffer composition

Do not exceed recommended running
conditions

Insufficient buffer

Fill inner and outer chambers to ensure
wells of gels are completely covered

Bands “smile” or “frown” within gel
lanes

Protein load too high

Load less protein

Sample or buffer preparation issues

Minimize salts, detergents, and solvents
in sample preparation and sample loading
buffers

Incorrect running conditions

Set correct voltage

Table 12 .1 . Troubleshooting electrophoresis and detection with Mini-PROTEAN

®

gels . For more troubleshooting tips,

refer to the relevant instrument instruction manuals or contact Technical Support.

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