Abstract
High-throughput, next-generation sequencing (NGS) offers a unique opportunity for in-depth characterization of adaptive immune receptor repertoires. Nevertheless, limitations and pitfalls exist in every step of both the experimental and the analytical procedure, leading to discrepancies in the literature and incomprehensive and/or altogether misleading results. Thus, standardization of protocols in NGS immunogenetics is urgently needed.
Here, we describe the experimental protocol that we developed for T-cell receptor beta chain (TRB) gene repertoire analysis in chronic lymphocytic leukemia, aiming to provide a reproducible and biologically meaningful output. Although optimized for TRBV-TRBD-TRBJ gene rearrangements, this protocol may be customized for other adaptive immune receptor sequences, as well.
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Financial Support
Supported in part by H2020 “AEGLE, An analytics framework for integrated and personalized healthcare services in Europe”, by the EU; “MEDGENET, Medical Genomics and Epigenomics Network” (No.692298) by the EU.
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Vlachonikola, E., Vardi, A., Stamatopoulos, K., Hadzidimitriou, A. (2019). High-Throughput Sequencing of the T-Cell Receptor Beta Chain Gene Repertoire in Chronic Lymphocytic Leukemia. In: Malek, S. (eds) Chronic Lymphocytic Leukemia. Methods in Molecular Biology, vol 1881. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8876-1_24
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DOI: https://doi.org/10.1007/978-1-4939-8876-1_24
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8875-4
Online ISBN: 978-1-4939-8876-1
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