Next Generation Sequencing
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loginThis book covers a wide range of various fields of research, with the common thread being NGS-related methods and applications, as well as some analysis and interpretation of the data obtained. The first two chapters focus on the highly dynamic processes of transla-tional and transcriptional profiling of a cell. In the first chapter, polysome and ribosome isolation by sucrose gradient is used to investigate translational activity, both approaches are described, and the information obtained from each is discussed. In the second chapter, isolation of cell nuclei and the nascent RNA transcripts therein allow a look at the nascent transcriptome of a cell. The third chapter focuses on a method to detect copy number alterations (CNAs) using whole genome amplification and low pass whole genome sequenc-ing. Chapters 4 and 5 touch on more targeted sequencing applications. The first uses small “bait” oligonucleotides to “fish” out long pieces of DNA (~2 Kb) from the genome com-bined with long read sequencing using the MinION (Oxford Nanopore, Inc.) allowing for the interrogation of these unknown flanking regions not contained in the baits. Chapter 5 deals with an extremely efficient and cost-effective method called “Hi-Plex” to characterize known polymorphic loci using a highly multiplexed amplicon-based approach to look for genetic variants.
