NGS – Next-generation sequencing

Developers

Companies: 454 Life Sciences, Illumina Inc., Life Technologies Corporation etc.

Description of the technology

Methods of the next generation sequencing (NGS) appeared thanks to the development of computer industry and technologies of production of microprocessors and digital data carriers.

NGS is a technic for determination the nucleotide sequence of DNA and RNA. Methods of the next generation sequencing allow to «read» simultaneously several genome regions, and this is the main characteristic which distinguishes NGS from earlier sequencing methods. NGS is realized through repeating cycles of the chain elongation using polymerase or multiple ligation of oligonucleotides.

Today, the next generation sequencing incorporates wide spectrum of methods which are basing of different principles and were developed more or less independently. The following technologies could be named herein: GWAS, WGBS, RRBS, De Novo sequencing, target sequencing, RNA-Seq, ChiP et al. Nowadays, these technologies are being renewed and modified very swift.

The next generation sequencing is considered to be a «second-generation» sequencing. Furthermore, platforms for the «third-generation» DNA sequencing have appeared up to date, and they grows rapidly. The third-generation sequencing differs from the second-generation one in that primary amplification of DNA is not further needed. DNA being examined is sequenced directly at the level of the single molecule without use of polymerases. This approach has certain advantages because it allows to prevent problems connected to accumulation of errors during DNA amplification. However, it should be said that none of the third-generation technologies has become widely distributed as yet.

Practical application

Invention and practical application of the next generation sequencing technologies have allowed to take to the next level such fields of science as genetics and molecular biology and stimulated formation of the personalized genome medicine.

Particularly, NGS technologies are used to study age-related changes in the human organism, investigate intra- and interspecific variations in the studies on polymorphisms of longevity-associated genes. ТNGS-method of transcriptome sequencing (RNA-seq) is used to study expression of gerontogenes. CHip-Seq technology finds application in the investigation of human epigenomes during aging.

Laboratories

• NGS platform, Institute of Genetics, Bern (Switzerland)
• 454 Life Sciences, Branford, Connecticut (USA), Basel (Switzerland)
• Illumina Inc., San Diego, California (USA)
• Life Technologies Corporation, Carlsbad, California (USA)
• Pacific Biosciences, California (USA)

Links

Publications

• Wheeler, David A., et al. «The complete genome of an individual by massively parallel DNA sequencing." nature 452.7189 (2008): 872–876.
• Ju, Young Seok, et al. «Extensive genomic and transcriptional diversity identified through massively parallel DNA and RNA sequencing of eighteen Korean individuals." Nature genetics 43.8 (2011): 745–752.
• de Magalhães, João Pedro, Caleb E. Finch, and Georges Janssens. «Next-generation sequencing in aging research: emerging applications, problems, pitfalls and possible solutions." Ageing research reviews 9.3 (2010): 315–323.
• Van Nieuwerburgh, Filip, et al. «Illumina mate-paired DNA sequencing-library preparation using Cre-Lox recombination." Nucleic acids research (2011): gkr1000.