E-Gel
™
Power Snap Electrophoresis System User Guide
31
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
•
Refer to the E-Gel
™
Sizing DNA Ladder migration pattern table to estimate target DNA
run time to the reference line.
•
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.
E-Gel
™
Sizing DNA Ladder migration pattern
Ladder
Fragment size
DNA amount (per 25 μL)
Migration time to
reference line
1,500 bp
1.5 ng
~19.5 min
1,200 bp
1.5 ng
~18.5 min
1,000 bp
6.0 ng
~17.5 min
900 bp
2.0 ng
~17 min
800 bp
2.0 ng
~16.5 min
700 bp
2.0 ng
~16 min
600 bp
2.0 ng
~15.5 min
500 bp
6.0 ng
~14.5 min
450 bp
2.0 ng
~14 min
400 bp
2.0 ng
~13.5 min
350 bp
2.0 ng
~13 min
300 bp
2.0 ng
~12.5 min
250 bp
2.0 ng
~11.5 min
200 bp
6.0 ng
~11 min
150 bp
2.0 ng
~10 min
125 bp
2.0 ng
~9.5 min
100 bp
2.0 ng
~9 min
75 bp
2.5 ng
~8.5 min
50 bp
2.5 ng
~8 min