Sanger and Next-Generation Sequencing of AAT

Valentina Barzon; Ilaria Ferrarotti; Stefania Ottaviani

doi:10.1007/978-1-0716-3605-3_6

Sanger and Next-Generation Sequencing of AAT

Methods Mol Biol. 2024:2750:57-67. doi: 10.1007/978-1-0716-3605-3_6.

Authors

Valentina Barzon¹, Ilaria Ferrarotti², Stefania Ottaviani³

Affiliations

¹ Department of Internal Medicine and Therapeutics, Pulmonology Unit, University of Pavia, Pavia, Italy.
² Centre for Diagnosis of Inherited Alpha-1 Antitrypsin Deficiency, UOC Pulmonology, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy. I.Ferrarotti@smatteo.pv.it.
³ Centre for Diagnosis of Inherited Alpha-1 Antitrypsin Deficiency, UOC Pulmonology, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy.

PMID: 38108967
DOI: 10.1007/978-1-0716-3605-3_6

Abstract

Sequencing of DNA is normally the final procedure carried out to determine the actual pathogenic variants when the techniques used for genotyping are unable to provide complete identification of both AAT alleles. Gene sequencing of complete SERPINA1 gene by using the Sanger method or next-generation sequencing (NGS) is crucial to enable correct diagnosis in patients with alpha1-antitrypsin deficiency caused by uncommon AAT variants.This protocol explains how to correctly sequence SERPINA1 gene both with Sanger method and NGS.

Keywords: Intron sequencing; Next-generation sequencing; Rare mutations; SERPINA1.

MeSH terms

Alleles
High-Throughput Nucleotide Sequencing*
Humans