Prepare Cassette
• Remove cassette from packaging and wipe off excess moisture. Verify correct cassette
type is used.
• Visually inspect buffer chamber and gel columns. From side view ensure buffer
chamber volume is more than ½ full. Inspect gel column for breakage, bubbles, or
gaps. Any lanes with these defects must be abandoned. Remaining lanes are still
functional.
• Inspect for air gaps in and behind elution wells by tilting cassette to the left (loading well
end). Tap on bench lightly to dislodge.
• Install cassette into tray of Pippin Prep with loading wells to the left.
• Remove sealing tape without tipping the cassette.
• Remove all buffer from elution well and replace with 40μl of electrophoresis buffer.
• Test electrophoretic current continuity:
• Close sliding lid on instrument.
• In the run screen select “Manual Mode” from the “Protocol Name” drop down menu.
• Press “Test” in controller on the Main Screen. Test takes 30 s. Finishing with “Pass” of
“Fail” message.
• If test fails—in the separation lane: lane unusable—no recovery.
• In the elution lane: Verify 40μl of running buffer is present in the elution well and
re-test. If it fails again, lane can be used for reference sample.
• Cover all collection wells with tape to minimize recover volume to ~50μl. Using razor
blade cut tape between elution wells so that.
• individual tape segments can be removed to minimize potential cross contamination.
Sample Loading
• Verify that sample wells are completely full. Top off wells with electrophoresis buffer if
necessary. Total well volume is 70μl.
• Remove 40μl of running buffer from the loading well leaving ~30μl in well.
• Load 40μl of pre-mixed Pippin Prep DNA Marker into the sample lane that has been
assigned for the reference. (Reference lane can be changed as needed.)
• Load 40μl of the supplemented sample into the appropriate lane loading well.
• Close lid.
• Select desired protocol from the “Protocol Name” drop down menu. (i.e.: TruSeq ChIP
250–300 bp.)
• Press “Start.”
Sample Collection
• When complete, remove elution well tape and transfer all samples from sample elution
well to a labeled microcentrifuge tube.
• Determine volume of collected sample. Volume can be from 40–70μl.
Appendix
205
• Remove cassette from packaging and wipe off excess moisture. Verify correct cassette
type is used.
• Visually inspect buffer chamber and gel columns. From side view ensure buffer
chamber volume is more than ½ full. Inspect gel column for breakage, bubbles, or
gaps. Any lanes with these defects must be abandoned. Remaining lanes are still
functional.
• Inspect for air gaps in and behind elution wells by tilting cassette to the left (loading well
end). Tap on bench lightly to dislodge.
• Install cassette into tray of Pippin Prep with loading wells to the left.
• Remove sealing tape without tipping the cassette.
• Remove all buffer from elution well and replace with 40μl of electrophoresis buffer.
• Test electrophoretic current continuity:
• Close sliding lid on instrument.
• In the run screen select “Manual Mode” from the “Protocol Name” drop down menu.
• Press “Test” in controller on the Main Screen. Test takes 30 s. Finishing with “Pass” of
“Fail” message.
• If test fails—in the separation lane: lane unusable—no recovery.
• In the elution lane: Verify 40μl of running buffer is present in the elution well and
re-test. If it fails again, lane can be used for reference sample.
• Cover all collection wells with tape to minimize recover volume to ~50μl. Using razor
blade cut tape between elution wells so that.
• individual tape segments can be removed to minimize potential cross contamination.
Sample Loading
• Verify that sample wells are completely full. Top off wells with electrophoresis buffer if
necessary. Total well volume is 70μl.
• Remove 40μl of running buffer from the loading well leaving ~30μl in well.
• Load 40μl of pre-mixed Pippin Prep DNA Marker into the sample lane that has been
assigned for the reference. (Reference lane can be changed as needed.)
• Load 40μl of the supplemented sample into the appropriate lane loading well.
• Close lid.
• Select desired protocol from the “Protocol Name” drop down menu. (i.e.: TruSeq ChIP
250–300 bp.)
• Press “Start.”
Sample Collection
• When complete, remove elution well tape and transfer all samples from sample elution
well to a labeled microcentrifuge tube.
• Determine volume of collected sample. Volume can be from 40–70μl.
Appendix
205