Description
The detailed protocol, outlining an easy, straightforward experimental workflow for generating a T-cell receptor α/β library can be found at: www.miltenyibiotec.com/applications/all-protocols/generation-of-tcr-alpha-and-beta-cdna-libraries-using-the-tcr-a-b-rna-profiling-kit-human or in the ‘Resources’ tab on this product page.
For molecular biology research use only. Not for diagnostic purposes.
Background information
Data analysis
A pre-set adapted specifically to the chemistry of the TCR α/β RNA Profiling Kit, human is available on Platforma, a no-code bioinformatics platform for NGS data. It can be accessed at https://platforma.bio. Platforma leverages MiXCR, the gold standard for immune repertoire analysis, and enables powerful downstream analysis such as repertoire diversity analysis, gene usage, rarefaction, and visualization, all without the need for coding.
Overview
The TCR α/β RNA Profiling Kit, human is designed to generate cDNA libraries of the alpha and beta T-cell receptor RNA for next-generation sequencing (NGS). This flexible solution can be initiated directly from sorted T cells, peripheral blood mononuclear cells (PBMCs), or RNA extracted from T cells or T cell containing tissues.
The complete protocol requires only 4–4.5 hours of hands-on time and has a total run time of 7.5 hours. It is designed for a wide range of input amounts, starting from as little as 0.1 ng of RNA (Figure 1). The product is adapted to support different sample qualities and generate high-quality results from FFPE-fixed samples (Figure 3). This is achieved by combining highly sensitive multiplex PCR with Unique Molecular Identifiers (UMIs). The kit's balanced chemistry ensures high reproducibility across experiments (Figure 2) and thus supports reliable and innovative molecular research.
