Cargando…

An open protocol for modeling T Cell Clonotype repertoires using TCRβ CDR3 sequences

T cell receptor repertoires can be profiled using next generation sequencing (NGS) to measure and monitor adaptive dynamical changes in response to disease and other perturbations. Genomic DNA-based bulk sequencing is cost-effective but necessitates multiplex target amplification using multiple prim...

Descripción completa

Detalles Bibliográficos
Autores principales: Gurun, Burcu, Horton, Wesley, Murugan, Dhaarini, Zhu, Biqing, Leyshock, Patrick, Kumar, Sushil, Byrne, Katelyn T., Vonderheide, Robert H., Margolin, Adam A., Mori, Motomi, Spellman, Paul T., Coussens, Lisa M., Speed, Terence P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10291816/
https://www.ncbi.nlm.nih.gov/pubmed/37365517
http://dx.doi.org/10.1186/s12864-023-09424-z
Descripción
Sumario:T cell receptor repertoires can be profiled using next generation sequencing (NGS) to measure and monitor adaptive dynamical changes in response to disease and other perturbations. Genomic DNA-based bulk sequencing is cost-effective but necessitates multiplex target amplification using multiple primer pairs with highly variable amplification efficiencies. Here, we utilize an equimolar primer mixture and propose a single statistical normalization step that efficiently corrects for amplification bias post sequencing. Using samples analyzed by both our open protocol and a commercial solution, we show high concordance between bulk clonality metrics. This approach is an inexpensive and open-source alternative to commercial solutions. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12864-023-09424-z.