The Third Revolution in Sequencing Technology

Trends Genet. 2018 Sep;34(9):666-681. doi: 10.1016/j.tig.2018.05.008. Epub 2018 Jun 22.


Forty years ago the advent of Sanger sequencing was revolutionary as it allowed complete genome sequences to be deciphered for the first time. A second revolution came when next-generation sequencing (NGS) technologies appeared, which made genome sequencing much cheaper and faster. However, NGS methods have several drawbacks and pitfalls, most notably their short reads. Recently, third-generation/long-read methods appeared, which can produce genome assemblies of unprecedented quality. Moreover, these technologies can directly detect epigenetic modifications on native DNA and allow whole-transcript sequencing without the need for assembly. This marks the third revolution in sequencing technology. Here we review and compare the various long-read methods. We discuss their applications and their respective strengths and weaknesses and provide future perspectives.

Keywords: long-read sequencing; nanopore sequencing; next-generation sequencing; single-molecule real-time sequencing; synthetic long-read sequencing; third-generation sequencing.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • DNA / analysis
  • DNA / genetics*
  • Exome Sequencing / methods
  • Exome Sequencing / trends*
  • Genome / genetics*
  • High-Throughput Nucleotide Sequencing / methods
  • High-Throughput Nucleotide Sequencing / trends*
  • Humans
  • Sequence Analysis, DNA / methods


  • DNA