How deeply should I sequence my Immunoverse™ TCR or BCR libraries?

How deeply you sequence your Immunoverse libraries depends on your application. Deeper sequencing will allow for the identification of more clonotypes; however, there is a plateau where the user will not see many additional clonotypes despite deeper sequencing. This plateau trends with library diversity, which is driven by input amount, input quality, immune cell content and other biological factors.

For dominant clone identification, the user may only need to sequence to 100,000 reads, while diversity or low abundance applications may require more input and deeper sequencing. A recommended starting point is 1.5 M reads with 400 ng of total RNA to capture most of the sample diversity in the sequencing data.

Panel Application RNA input recommendations* Sequencing read depth
TCR B/G Clonality or dominant clones 25-400 ng 250,000
TCR A/D Clonality or dominant clones 25-400 ng 250,000
TCR All Chains Clonality or dominant clones 25-400 ng 500,000
TCR B/G Thorough characterization of repertoire or rare clonotype identification 400-2000 ng 1.5 M
TCR A/D Thorough characterization of repertoire or rare clonotype identification 400-2000 ng 1.5 M
TCR All Chains Thorough characterization of repertoire or rare clonotype identification 400-2000 ng 3 M
BCR IGH Clonality or dominant clones 100-400 ng 250,000
BCR IGK/L Clonality or dominant clones 100-400 ng 250,000
BCR All Chains Clonality or dominant clones 100-400 ng 500,000
BCR IGH Thorough characterization of repertoire or rare clonotype identification 400-6000 ng 1.5 M
BCR IGK/L Thorough characterization of repertoire or rare clonotype identification 400-6000 ng 1.5 M
BCR All Chains Thorough characterization of repertoire or rare clonotype identification 400-6000 ng 3 M

* Depending on sample T and B cell content. See the following resources:

The rarefaction analysis below was derived from Immunoverse TCR libraries generated from varying amounts of high-quality input from a PBL sample and illustrates the relationship between input amounts, sequencing depths and clonotype count for a diverse sample type.

Sequencing depth, input amount, and library diversity influence the amount of clonotypes that can be identified by Immunoverse assays.


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For Research Use Only. Not for use in diagnostic procedures. For Research Use Only. Not for use in diagnostic procedures.