Library Prep Validation
Last updated
Last updated
This single-step protocol models the library prep required to produce libraries tagged with index sequences that are ready for the various sequencing workflow.
In Lab View, locate the Library Prep Validation protocol.
Samples are queued for the Library Prep Validation step.
Add the samples to the Ice Bucket and select View Ice Bucket.
On the Ice Bucket screen, select the label groups (e.g., TruSeq HT Adapters v2 (D7-D5)) to use.
[Optional] Manually create reagent labels (indexes) in the Configuration screen under Consumables > Labels.
For more information on how to create reagent label group, refer to the Add and Configure Labels and Label Groups section in the Clarity LIMS (Clarity & LabLink Reference Guide) documentation.
Select Begin Work.
On the Placement screen, perform the following actions:
In the Placed Samples area in the Container field, scan or enter the barcode of the destination container.
Select the samples from the Samples to be Placed area, then drag them over to the container wells on the right.
Select Add Labels.
On the Add Labels screen, the Labels to Add list is populated with labels from the selected label group.
Select labels on the left, and drag them over to the container on the right to assign them to placed samples.
Select Record Details.
On the Record Details screen, perform the following actions:
In the Sample Details table, enter values for Molarity (nM) for all the individual sample. Enter values individually or use the copy/paste functionality to enter multiple values from a spreadsheet.
Molarity (nM) values are used in Calculate Volume automation script in some workflow steps. If Molarity (nM) values are not entered, the Calculate Volume automation script does not work properly.
The Sequencing Instrument column indicates the instrument to be used for the run. Select the instrument to be routed.
Populate the fields in this column using one of the following methods:
To populate fields individually for each sample, select from the drop-down list in each row.
To populate fields all at one time, use the Copy and Paste buttons to enter multiple values from a spreadsheet (refer to the following note).
Alternatively, select inside the table and press Ctrl + A to select all rows. At the top of the table, select Sequencing Instrument, then select the instrument to be routed from the adjacent drop-down list. Select Apply.
If working with many samples, copy and paste to insert multiple values from a spreadsheet as follows.
Select the Copy button to copy the sample information from Clarity LIMS to the clipboard.
Open a blank Excel document and paste the contents of the clipboard into the spreadsheet.
In the spreadsheet, enter the Molarity (nM) and Sequencing Instrument values. Select the entire spreadsheet, including the headers, (Ctrl + A) and copy (Ctrl + C) the information to the clipboard.
In Clarity LIMS, select the Paste button. Paste the contents of the clipboard (Ctrl + V) into the Update Table window and then select Update to paste into the Sample Details table.
Select Next Steps.
This selection triggers the Next Steps automation, which sets the value of the next step (for all samples) to Remove from workflow. The Routing Script automation expects this value and requires it to advance samples to the next step successfully.
On the Assign Next Steps screen in the Samples table, the Next Step for all samples is prepopulated with Remove from workflow.
Do not change this value. If Next Step is not set to Remove from workflow, the routing script is not able to route samples correctly.
Select Finish Step.
On exit from the step, the Routing Script automation is triggered. This automation assigns samples to the first step of the sequencing workflow of the selected instrument.
