High-Throughput Contiguous Full-Length Next-Generation Sequencing of HLA Class I and II Genes from 96 Donors in a Single MiSeq Run

Methods Mol Biol. 2018:1802:89-100. doi: 10.1007/978-1-4939-8546-3_6.

Abstract

The human leukocyte antigen (HLA) genes regulate and drive the immune system, and are among the most polymorphic loci in the human genome. HLA diversity is known to play an important role in transplantation and disease association studies. There are multiple approaches to DNA-based HLA genotyping and recent advances in next-generation sequencing (NGS) technologies have facilitated the development of whole gene sequencing methods. We describe an accurate, efficient, scalable, and cost-effective approach to contiguously amplify and sequence full-length genes of six HLA class I and II loci from 96 individuals on a single Illumina MiSeq run.

Keywords: Full-length amplification; HLA; HLA genotyping; Illumina; MIT-NGS; MiSeq; NGS; Next-generation sequencing.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Gene Library
  • Genetic Loci
  • High-Throughput Nucleotide Sequencing / methods*
  • Histocompatibility Antigens Class I / genetics*
  • Histocompatibility Antigens Class II / genetics*
  • Humans
  • Polymerase Chain Reaction
  • Tissue Donors*

Substances

  • Histocompatibility Antigens Class I
  • Histocompatibility Antigens Class II