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PippinHT Operations Manual
2-3
460005 Rev G
2.2 Improving Sample Yield
Users should expect an intrinsic yield of between 50-80% of the sample input. This is
based on product validation studies in which restriction fragments have been run and
collected, and the respective DNA amounts compared.
Yield can be affected by DNA becoming lightly bound to the low molecular weight cut-
off membrane in the elution well. At the end of the run, the PippinHT will briefly
reverse field (for 5 seconds) in every elution channel. This improves sample yield by
up to 20%.
Note:
The reverse field feature may be disabled in the Systems Option screen.
2.3 Improving Sample Yield for HMW DNA (High-Pass
Protocols)
Recovering high molecular weight DNA from High-Pass protocols tend to have lower
yields than low molecular weight size selection. The following additional steps are
highly recommended to improve product yield:
1. Wait
at least 45 min
after termination of the run before removing eluted DNA (it is
OK to leave samples in instrument for hours, i.e. for an overnight run).
2. Rinse the elution well with Tween solution. Tween solution (0.1% Tween 20 in
electrophoresis buffer) is provided by Sage Science with cassette kits used with
HMW collections.
After removing the initial eluted volume:
•
Remove the eluted sample from elution well at the end of the run
•
Add 30
l of Tween solution (supplied with the gel cassettes) to the elution well
•
Wait 1 minute
•
Remove solution and pool with the original extracted sample, or process
separately
.
