@inbook{f7a372c1b4a5459cbb18b23081240d4a,
title = "The Experimental Procedure of Somatic Hypermutation Analysis in Splenic B Cells During Primary Response",
abstract = "Somatic hypermutation (SHM) is a critical process in adaptive immunity, enabling the generation of high-affinity antibodies through targeted mutations in immunoglobulin variable (IgV) regions. Here, we provide a comprehensive workflow combining immunization, molecular biology, and bioinformatics to investigate SHM mechanisms and outcomes. This is a general protocol for studying SHM at the VH186.2 region in C57BL/6 mice. While this method can be applied broadly, this chapter will detail the protocol used to test the effects of exonuclease 1(EXO1) knock-in mutation (Exo1D173A, or Exo1DA) or knock-out (KO) on hypermutation post-immunization by immunizing age-matched mice with NP(33)-CGG on alum. We start by immunizing and sacrificing the animals to obtain spleens for RNA extraction. We then create cDNA libraries and investigate VH186.2 region mutation to analyze SHM.",
keywords = "Activation-induced cytidine deaminase, Adaptive immunity, AID, B-cell receptor, BCR, Next-generation sequencing, Oncogenesis, SHM, Somatic hypermutation",
author = "Seth Greene and Alyssa Fennig-Victor and Yongwei Zhang and Shanzhi Wang",
note = "Publisher Copyright: {\textcopyright} The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2026.",
year = "2026",
doi = "10.1007/978-1-0716-4901-5\_36",
language = "English (US)",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "493--499",
booktitle = "Methods in Molecular Biology",
}