Determining immunoglobulin-specific B cell receptor repertoire of murine splenocytes by next-generation sequencing

STAR Protocols

Simon Schäfer¹  Falk Nimmerjahn¹  Anja Werner²  Thomas H. Winkler³  Nina Gleußner³ 

[1] Corresponding author;Division of Genetics, Department of Biology, Friedrich-Alexander-University Erlangen-Nürnberg (FAU), Erwin-Rommelstr. 3, 91058 Erlangen, Germany;Division of Genetics, Department of Biology, Friedrich-Alexander-University Erlangen-Nürnberg (FAU), Glückstrasse 6, 91054 Erlangen, Germany;

关键词: Bioinformatics;    Sequence analysis;    Cell Biology;    Cell isolation;    Sequencing;    Immunology;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: High-throughput antibody repertoire analysis via next-generation sequencing is a key method in understanding humoral immunity. Errors introduced during library preparation and sequencing can be corrected with molecular amplification fingerprinting using unique molecular identifiers. Here, we provide a step-by-step protocol for laboratory and bioinformatic workflows to perform sequencing in murine cells with isotype-specific primers, obtaining total and isotype-specific B cell receptor repertoires. This enables the examination of isotype-dependent immune responses and improves the understanding of underlying mechanisms in humoral immunity.For complete details on the use and execution of this protocol, please refer to Khan et al. (2016).

【 授权许可】

Unknown