Next Generation Sequencing

This volume covers a wide range of various fields of research, with the common thread being Next Generation Sequencing (NGS) related methods and applications, as well as analysis and interpretation of the data obtained. Chapters guide readers through the highly dynamic processes of translational and transcriptional profiling of a cell, method to detect copy number alterations (CNAs), targeted sequencing applications, method called "Hi-Plex" to characterize known polymorphic loci, single-cell of DNA or RNA, identify and characterize rare circulating CD4 T cells, and computational pipeline for RNAseq analysis. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Next Generation Sequencing: Methodsand Protocols aims to be useful and informative for further study into this vital field.

An Integrated Polysome Profiling and Ribosome Profiling Method to Investigate in vivo Translatome. - Measuring Nascent Transcripts by Nascent-seq. - Genome-wide Copy Number Alteration Detection in Preimplantation Genetic Diagnosis. - Multiplexed Targeted Sequencing for Oxford Nanopore MinION; A Detailed Library Preparation Procedure. - Hi-Plex for Simple, Accurate, and Cost-effective Amplicon-based Targeted DNA Sequencing. - ClickSeq: Replacing Fragmentation and Enzymatic Ligation with Click-Chemistry to Prevent Sequence Chimeras. - Genome-wide Analysis of DNA Methylation in Single Cells Using a Post-bisulfite Adapter Tagging Approach. - Sequencing of Genomes from Environmental Single Cells. - SNP Discovery from Single and Multiplex Genome Assemblies of Non-model Organisms. - CleanTag Adapters Improve Small RNA Next-Generation Sequencing Library Preparation by Reducing Adapter Dimers. - Sampling, Extraction, and High-Throughput Sequencing Methods for Environmental Microbial and Viral Communities. - A Bloody Primer Analysis of RNA-Seq from Tissue Admixtures. - Next-Generation Sequencing of Genome-wide CRISPR Screens. - Gene Profiling and T Cell Receptor Sequencing from Antigen-specific CD4 T Cells. - Investigate Global Chromosomal Interaction by Hi-C in Human Naïve CD4 T Cells. - Primer Extension, Capture, and On-bead cDNA Ligation: An Efficient RNAseq Library Prep Method for Determining Reverse Transcription Termination Sites.