Troubleshooting - Thermo Scientific Invitrogen E-Gel G8100 User Manual

Troubleshooting

For common E-Gel ™ troubleshooting guidelines refer to troubleshooting guide (see page 35).
Observation
Poor resolution or
smearing of bands
Low yield
Target DNA band cannot
be seen
DNA band passed the
recovery gel
DNA migration exhibits
smiley effect
28
Cause
Sample is overloaded
High salt concentration
Total sample volume is too low
or too high
Loading wells were not pre-
filled with deionized water prior
to loading the sample
Samples were not prepared
properly
Incorrect loading volume
chosen
Recovery wells were not filled
with water prior to elution
DNA band passed the recovery
gel
DNA band amount is too high
High ambient light or low
sample amount
Selected protocol time was too
long
Extended gel run time or aged
gels used or incorrect loading
conditions
Recommended action
Do not load more than 800 ng of DNA in a single
lane
Dilute your samples 2- to 30-fold
Load recommended sample volume of 25 μL per
lane.
Fill all gel wells with 50 μL of deionized water prior
to loading any sample or a ladder.
Prepare up to 25 μL of sample in 1X concentration
of 10X Sample Loading Buffer.
Load up to 25 μL of prepared sample per well
Once target fragment reaches reference line, pause
the run and fill all recover wells with deionized
water.
Carefully observe the band migration into the
recovery well. Minimize ambient light or perform
the workflow in dark room.
Collect DNA from the well in two or more fractions.
Be sure to load the recommended DNA amount.
Perform the workflow in dark room environment to
minimize ambient lights;
confirm sample concentration prior to loading
Choose the Reverse E-Gel program to run the band
backwards into the collection well
Do not run gels longer than 40 minutes. Use fresh
gel. Follow sample loading recommendations.
E-Gel Power Snap Electrophoresis System User Guide
™