Generally, more input will allow for identification of more clonotypes at any level of sequencing. The TCR assay will tolerate between 25 ng - 2 ug of high quality input. Inputs between 100 ng and up to 6 ug of high quality diverse PBL RNA have been tolerated in the BCR assay. For diversity applications, we recommend using as much available input as possible to increase the total number of clonotypes identified. For dominant clonotyping applications, less input can be used but we recommend starting with 200-400ng if possible.
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.
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