2020 Short Course - Essential Fundamentals - Next Generation Sequencing From The Beginning-NGS Technologies

Video Transcription

Video Summary

In this tutorial on next-generation sequencing technologies, Elaine Martis from Ohio State University and Nationwide Children's Hospital explains the basics of how next-generation sequencing works and its significance in biomedical research. She discusses the transformational impact of next-generation sequencing on biomedical inquiry and provides a comparison of traditional Sanger sequencing versus next-generation sequencing methodologies. Martis explains the steps involved in next-generation sequencing, including library construction, amplification, and sequencing reactions. She highlights the advantages of next-generation sequencing, such as its ability to couple sequencing chemistry with detection, resulting in higher throughput and simplicity. Martis also discusses the input DNA requirements for library construction and the need for DNA isolation and evaluation of quantity and quality. She delves into the different sequencing platforms, focusing on Illumina sequencing (sequencing by synthesis) and Ion Torrent sequencing (sequencing by pH sensing). Martis briefly discusses the use of hybrid capture-based approaches to subset the genome and explains the importance of alignment and variant calling in the analysis of next-generation sequencing data. She concludes by emphasizing the need for careful consideration and validation of approaches in the clinical space, as well as the importance of visualizing data and characterizing variants for clinical sign-out reports.

Asset Subtitle

Elaine Mardis

Keywords

next-generation sequencing

NGS

sequencing technologies

biomedical research

Sanger sequencing

library construction

amplification

sequencing reactions

Illumina sequencing

Ion Torrent sequencing

hybrid capture-based approaches