Step 3: Dilute and Denature (NovaSeq X Series Sequencing v1.0)
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1.
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In Lab View, locate the NovaSeq X Series Sequencing v1.0 protocol. The pool of samples queued for the Dilute and Denature (NovaSeq X Series Sequencing v1.0) displays. |
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2.
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Add the pool to the Ice Bucket and select View Ice Bucket. |
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3.
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[Optional] On the Ice Bucket screen, set the number of derivatives to create (placed into the library tube strip) and select Begin Work. |
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On entry to the step, the Validate Inputs Flowcell Type and Register Step Started automation is triggered. This automation makes sure that the configured flow cell type is valid. The automation also registers the start time of the step by publishing messages to CLPA through the ICA LINK service. The portion of the script used for start time registration is used only for CLPA support. |
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On entry to the Record Details screen, the Calculate Volumes automation is triggered. This automation sets the following values: |
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The automation also generates the calculation file (CSV) and attaches it to the step. This file contains information about the volume of RSB, NaOH, and TT2 to add per working pool. |
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4.
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On the Record Details screen, the Reagent Lot Tracking section tracks the NaOH, Resuspension Buffer, and TT2 reagents used in the step. To add and activate reagent lots, do as follows. |
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a.
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In a new browser tab, open an additional instance of Clarity LIMS. |
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b.
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Navigate to the Consumables configuration screen. |
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c.
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Select one of the reagent kits previously listed (eg, NaOH) and select New Lot. |
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d.
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Enter the lot details. |
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f.
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Set the Status of Reagent Lot toggle switch to Active. |
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g.
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Repeat this process to add and activate the remaining reagent lots. |
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h.
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Return to your original working browser tab and refresh the page. |
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5.
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On the Record Details screen, perform the following actions: |
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a.
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In the Reagent Lot Tracking section, select from the active lots displayed in each drop-down list. |
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b.
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In the Sample Details table, make sure that the BP aliquot, RSB, NaOH, and TT2 reagent volume values are populated. The script sets these values and they cannot be edited. |
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c.
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[Optional] In the Sample Details table, select the pool icon to view details of the working pool composition. |
The working pool number is appended to the bulk pool name so that you can identify which working pools are derived from the same bulk pool.
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6.
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In the Files area, select the Calculation File (CSV) to open it and view details on the RSB, NaOH, and TT2 volumes to add per working pool. |
