Collect Dna Fragment; Guidelines For Estimating Run Time - Thermo Scientific Invitrogen E-Gel G8100 User Manual

Collect DNA fragment

1. Resume the run and carefully observe as
the reference band enters the recovery well.
Important: See NGS library size selection
reference to determine when to collect
samples of specific target library length.
2. Stop the gel and recover the sample with a
pipette. Avoid piercing the agarose.
Some residual DNA will remain visible in
the well due to migration into the agarose
at the bottom of the well.
3. Proceed with downstream NGS workflow.
4. (Optional) Use the Reverse E-Gel protocol
if the band of interest passes the recovery
well (see page 20).

Guidelines for estimating run time

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E-Gel Sizing DNA Ladder migration pattern
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Ladder
E-G el Power Snap Electroph oresis System User Guide
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Refer to the E-Gel Sizing DNA Ladder migration pattern table to estimate target DNA
run time to the reference line.
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The E-Gel DNA Sizing Ladder is also used as a size reference marker. Refer to the NGS
library size selection reference to estimate run time from the reference line to the
collection well.
The run times indicated in the table are estimates. Monitor your gel in real time during
the run to ensure the sample does not pass the recovery well.
Identically sized bands in different wells may migrate differently.
DNA fragment size, amount, and salt content can affect migration rates.
Fragment size
1,500 bp
1,200 bp
1,000 bp
900 bp
800 bp
700 bp
600 bp
500 bp
450 bp
400 bp
350 bp
300 bp
250 bp
200 bp
150 bp
125 bp
100 bp
75 bp
50 bp
DNA amount (per 25 μL)
1.5 ng
1.5 ng
6.0 ng
2.0 ng
2.0 ng
2.0 ng
2.0 ng
6.0 ng
2.0 ng
2.0 ng
2.0 ng
2.0 ng
2.0 ng
6.0 ng
2.0 ng
2.0 ng
2.0 ng
2.5 ng
2.5 ng
Migration time to
reference line
~19.5 min
~18.5 min
~17.5 min
~17 min
~16.5 min
~16 min
~15.5 min
~14.5 min
~14 min
~13.5 min
~13 min
~12.5 min
~11.5 min
~11 min
~10 min
~9.5 min
~9 min
~8.5 min
~8 min
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