Using synthetic templates to design an unbiased multiplex PCR assay

TLDR: A multiplex PCR with a mixture of primers targeting the rearranged variable and joining segments to capture receptor diversity is applied to a multiplex T cell receptor gamma sequencing assay and can be extended to any adaptive immune locus.

Abstract: Immunosequencing enables cost-effective sequencing of repertoires of immune cells, but it often suffers from amplification biases when attempting cell quantification. Here, the authors present a powerful multiplex PCR assay that allows for quantitative and unbiased analysis of frequency of different T cell receptors.