This guide aims at getting you up and running quickly with Geneious Biologics and includes the following topics:
- Adding users
- Workspace and Folders
- Folder sharing
- Importing and uploading files
- Running an NGS analysis with the Antibody annotator
- Running a Sanger Analysis
- Visualizing results
- Barcodes, UMIs, Single Clone Analysis and 10X
- Need more help?
More detailed articles can be found in this knowledge base, and several are directly linked within this guide.
To add a new user into your organisation, click Users in the sidebar and click New in the main page. Only administrators can manage users.
Fill in the appropriate details and click Create User.
A Geneious account activation email will be sent to the email address which contains instructions on password setup and account activation.
Note that only admin accounts have administrative rights to create user accounts, name schemes, scaffolds and primer sets.
Workspace and Folders
Folders (folder icon in the folder tree) may contain any mixture of documents, such as nucleotide sequences, protein sequences, sequence alignments, and trees.
To create a folder, hover over Shared workspace, click the menu button (3 vertical dots) and click New folder in the popup menu.
Once you’ve created a folder, you can create more Folders within this folder by hovering your mouse over the folder and clicking the menu button.
Files can be moved from one folder to another using the "Move" icon at the top of the document table.
By default, any new folder is not shared until you set up the sharing options. To share folders, select Manage Sharing in the popup menu for folder, which can be accessed by clicking on the menu button (vertical ellipsis).
You can set the access level for all members of your organization as well as add new members by typing your colleagues’ name or email address into the field and select the user you wish to add.
Note that you can only manage sharing of your top level folders. You can find out more about sharing folders here.
Importing and uploading files
Once you have created a folder, files can be uploaded using the Upload button or via drag-and-drop.
Running an NGS analysis with the Antibody annotator
Set and merge paired reads
To pair reads, select a fastq file or multiple fastq files, click Pre-processing and click Set & merge paired read in the dropdown.
To pair corresponding reads, select the appropriate settings for the data and click Run. In this example, the data are paired and merged.
Once the reads are merged, a new document containing the merged reads will be generated (as well as one contained the unmarked ones). To run an NGS analysis with the Antibody annotator, select the merged data, click Annotation and select Antibody annotator in the dropdown.
Once the appropriate settings have been selected, click Run to start the analysis. You can find out more about the options available here or by clicking the Help button on the bottom right corner of the Antibody annotator popup.
The Antibody annotator will output a result document with the words Annotated & Clustered appended to the end of your input file name.
Running a Sanger Analysis with the Antibody Annotator
Sanger analysis in Geneious Biologics is much the same process as the NGS workflow described above. The main differences are the preprocessing steps used to clean your data prior to running Antibody Annotator, and how you choose to continue your analysis after annotation.
Preprocessing steps currently used on Sanger sequences include: Batch Assembly, Associate Heavy/Light Chains, Find Heterozygotes , and Batch Rename. These steps can be used alone or in combination.
Note that Batch Assembly and Associate Heavy/Light Chains may require that sequences have consistent naming schemes so that they can be matched correctly. This can be achieved with the "Batch rename" operation, which renames sequences by replacing or removing existing sections of the name.
If you wish to see parts of your sequence names (such as Well ID) in the annotation Result "All Sequences" table, this can be achieved by setting up a File Naming Scheme. File naming schemes are particularly useful for creating columns to use when adding extra experimental data to your sequences. For more information about adding assay data to your results, see here. You can also add labels and free-form text notes to your sequences after annotation.
Once the analysis is finished, you can visualise the results by selecting the Antibody annotator output file. The Pipeline Report gives an overview of the results which also consist of details on the Heavy and Light CDR3 diversity alongside VJ association in the form of heatmaps. You can export this report as a pdf file by clicking Export to PDF.
To view individual reads, select the NGS analysis output file, click the Sequences tab and select the reads that you would like to have a closer look at.
You can also group the reads by Heavy or Light CDR3 by selecting Heavy CDR3 in the Group by dropdown.
The NGS result table can be filtered by specifying a valid SQL condition using column headers as the fields to filter on.
To search for sequences that meet the condition of having both heavy and light chains with a heavy CDR3 motif of ‘AKNQGYVFGNWFFDH’, you can either right click an appropriate cell with the selected conditions to filter on or input a syntax as such: chain = 'Both' AND heavy_cdr3 = 'AKNQGYVFGNWFFDH'
Once you have specify your syntax, hit Enter or click Filter. The Sequences table will refresh and only sequences that fit the specified conditions will be shown in the results table. You can learn more about NGS data filtering and SQL syntax used in Geneious Biologics here.
To have a quick overview of the cluster lengths, click the Graphs tab and select Cluster lengths in the dropdown.
To have a quick overview of the lengths of the heavy CDR3 cluster, select Heavy CDR3 in the Show dropdown.
The graph in view can be exported as an image file by clicking Export Graph.
Notice that all export operations from the graphs are automatically downloaded, while other export operations need the user to download the exported document from the jobs table (see screenshot below).
Barcodes, UMIs and Single Clone Analysis
These are new features. We are currently writing articles to address the topics of how to collapse Unique Molecular Identifiers, and assemble sequences using Barcodes. We are also writing up help for Single Clone/Single Cell in a Well and 10X Antibody workflows.
Feel free to contact us directly with questions in the meantime.
Need more help?
For further questions, please check many others articles in our knowledge base or submit a help request by navigating to the help page, and then clicking Contact Support.