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*WarningLog*.tsv lists warnings that occurred during a run.
Rehybridization Workflow
A rehybridization run might be necessary if metrics generated during the first few cycles show intensities
below 2500. Some low-diversity libraries can show intensities below 1000, which is expected and cannot be
resolved with rehybridization.
NOTE
The End Run command is final. The run cannot be resumed, run consumables cannot be reused, and
sequencing data from the run are not saved.
When you end a run and save the flow cell, the software performs the following steps before the run ends:
u
Places the flow cell in a safe state.
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Unlocks the flow cell RFID for a later run.
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Assigns a rehybridization expiration date to the flow cell.
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Writes the run logs for completed cycles. A delay is normal.
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Bypasses the automatic post-run wash.
When you start a rehybridization run, the software performs the following steps to perform the run:
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Creates a run folder based on a unique run name.
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Checks that the flow cell rehybridization date has not expired.
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Primes reagents. A delay is normal.
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Skips the clustering step.
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Removes the previous Read 1 primer.
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Hybridizes a fresh Read 1 primer.
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Continues through Read 1 and the remainder of the run based on specified run parameters.
Points to End a Run for Rehybridization
Later rehybridization is possible only if you end the run at the following points:
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After cycle 5—Intensities appear after template registration, which requires the first 5 cycles of
sequencing. Although it is safe to end a run after cycle 1, ending after cycle 5 is recommended. Do not
end a run during cluster generation.
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Read 1 or Index 1 Read—End the run
before
paired-end resynthesis begins. The flow cell cannot be
saved for later rehybridization after paired-end resynthesis begins.
Required Consumables
A rehybridization run requires a new MiniSeq reagent cartridge regardless of when the run was stopped.
End the Current Run
1
Select End Run. When prompted to confirm the command, select Yes.
2
When prompted to save the flow cell, select Yes. Note the expiration date for rehybridization.
3
Remove the saved flow cell and set aside at 2°C to 8°C until you are ready to set up the rehybridization
run.
Document # 1000000002695 v02 Material # 20014309
For Research Use Only. Not for use in diagnostic procedures.
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MiniSeq System Guide