Tracking B-cell repertoires and clonal histories in normal and malignant lymphocytes
Tracking B-cell repertoires and clonal histories in normal and malignant lymphocytes
Methods for tracking B-cell repertoires and clonal history in normal and malignant B-cells based on immunoglobulin variable region (IGV) gene analysis have developed rapidly with the advent of massive parallel next-generation sequencing (mpNGS) protocols. mpNGS permits a depth of analysis of IGV genes not hitherto feasible, and presents challenges of bioinformatics analysis, which can be readily met by current pipelines. This strategy offers a potential resolution of B-cell usage at a depth that may capture fully the natural state, in a given biological setting. Conventional methods based on RT-PCR amplification and Sanger sequencing are also available where mpNGS is not accessible. Each method offers distinct advantages. Conventional methods for IGV gene sequencing are readily adaptable to most laboratories and provide an ease of analysis to capture salient features of B-cell use. This chapter describes two methods in detail for analysis of IGV genes, mpNGS and conventional RT-PCR with Sanger sequencing.
281-301
Weston Bell, Nicola
c99c8c28-519f-4c3e-bd8c-679995a0472e
Cowan, Graeme
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Sahota, Surinder
66c1457f-cba2-4c49-9c8c-fcee0748b6b8
2017
Weston Bell, Nicola
c99c8c28-519f-4c3e-bd8c-679995a0472e
Cowan, Graeme
7ff34a9b-f730-4a1e-8f32-634f709b63fd
Sahota, Surinder
66c1457f-cba2-4c49-9c8c-fcee0748b6b8
Weston Bell, Nicola, Cowan, Graeme and Sahota, Surinder
(2017)
Tracking B-cell repertoires and clonal histories in normal and malignant lymphocytes.
In,
Calado, D.
(ed.)
Germinal Centers.
(Methods in Molecular Biology, 1623)
New York, NY.
Humana Press, .
(doi:10.1007/978-1-4939-7095-7_21).
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Book Section
Abstract
Methods for tracking B-cell repertoires and clonal history in normal and malignant B-cells based on immunoglobulin variable region (IGV) gene analysis have developed rapidly with the advent of massive parallel next-generation sequencing (mpNGS) protocols. mpNGS permits a depth of analysis of IGV genes not hitherto feasible, and presents challenges of bioinformatics analysis, which can be readily met by current pipelines. This strategy offers a potential resolution of B-cell usage at a depth that may capture fully the natural state, in a given biological setting. Conventional methods based on RT-PCR amplification and Sanger sequencing are also available where mpNGS is not accessible. Each method offers distinct advantages. Conventional methods for IGV gene sequencing are readily adaptable to most laboratories and provide an ease of analysis to capture salient features of B-cell use. This chapter describes two methods in detail for analysis of IGV genes, mpNGS and conventional RT-PCR with Sanger sequencing.
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e-pub ahead of print date: 7 June 2017
Published date: 2017
Identifiers
Local EPrints ID: 417877
URI: http://eprints.soton.ac.uk/id/eprint/417877
PURE UUID: 370c1be4-5fe1-4843-88df-8762f42fa7ad
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Date deposited: 16 Feb 2018 17:30
Last modified: 15 Mar 2024 18:29
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Contributors
Author:
Nicola Weston Bell
Author:
Graeme Cowan
Author:
Surinder Sahota
Editor:
D. Calado
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