Introduces readers to core algorithmic techniques for next-generation sequencing (NGS) data analysis and discusses a wide range of computational techniques and applications This book provides an in-depth survey of some of the recent developments in NGS and discusses mathematical and computational challenges in various application areas of NGS technologies. The 18 chapters featured in this book have been authored by bioinformatics experts and represent the latest work in leading labs actively contributing to the fast-growing field of NGS. The book is divided into four parts: Part I focuses on computing and experimental infrastructure for NGS analysis, including chapters on cloud computing, modular pipelines for metabolic pathway reconstruction, pooling strategies for massive viral sequencing, and high-fidelity sequencing protocols. Part II concentrates on analysis of DNA sequencing data, covering the classic scaffolding problem, detection of genomic variants, including insertions and deletions, and analysis of DNA methylation sequencing data. Part III is devoted to analysis of RNA-seq data. This part discusses algorithms and compares software tools for transcriptome assembly along with methods for detection of alternative splicing and tools for transcriptome quantification and differential expression analysis. Part IV explores computational tools for NGS applications in microbiomics, including a discussion on error correction of NGS reads from viral populations, methods for viral quasispecies reconstruction, and a survey of state-of-the-art methods and future trends in microbiome analysis. Computational Methods for Next Generation Sequencing Data Analysis: Reviews computational techniques such as new combinatorial optimization methods, data structures, high performance computing, machine learning, and inference algorithms Discusses the mathematical and computational challenges in NGS technologies Covers NGS error correction, de novo genome transcriptome assembly, variant detection from NGS reads, and more This text is a reference for biomedical professionals interested in expanding their knowledge of computational techniques for NGS data analysis. The book is also useful for graduate and post-graduate students in bioinformatics.

Recently, next generation sequencing (NGS) technology has emerged as a powerful approach and dramatically transformed biomedical research in an unprecedented scale. NGS is expected to replace the traditional hybridization-based microarray technology because of its affordable cost and high digital resolution. Although NGS has significantly extended the ability to study the human genome and to better understand the biology of genomes, the new technology has required profound changes to the data analysis. There is a substantial need for computational methods that allow a convenient analysis of these overwhelmingly high-throughput data sets and address an increasing number of compelling biological questions which are now approachable by NGS technology. This dissertation focuses on the development of computational methods for NGS data analyses. First, two methods are developed and implemented for detecting variants in analysis of individual or pooled DNA sequencing data. SNVer formulates variant calling as a hypothesis testing problem and employs a binomial-binomial model to test the significance of observed allele frequency by taking account of sequencing error. SNVerGUI is a GUI-based desktop tool that is built upon the SNVer model to facilitate the main users of NGS data, such as biologists, geneticists and clinicians who often lack of the programming expertise. Second, collapsing singletons strategy is explored for associating rare variants in a DNA sequencing study. Specifically, a gene-based genome-wide scan based on singleton collapsing is performed to analyze a whole genome sequencing data set, suggesting that collapsing singletons may boost signals for association studies of rare variants in sequencing study. Third, two approaches are proposed to address the 3'UTR switching problem. PolyASeeker is a novel bioinformatics pipeline for identifying polyadenylation cleavage sites from RNA sequencing data, which helps to enhance the knowledge of alternative polyadenylation mechanisms and their roles in gene regulation. A change-point model based on a likelihood ratio test is also proposed to solve such problem in analysis of RNA sequencing data. To date, this is the first method for detecting 3'UTR switching without relying on any prior knowledge of polyadenylation cleavage sites.

A Practical Guide to the Highly Dynamic Area of Massively Parallel SequencingThe development of genome and transcriptome sequencing technologies has led to a paradigm shift in life science research and disease diagnosis and prevention. Scientists are now able to see how human diseases and phenotypic changes are connected to DNA mutation, polymorphi

Advancements in next-generation sequencing (NGS) technology have enabled the rapid growth and availability of large quantities of DNA and RNA sequences. These sequences from both model and non-model organisms can now be acquired at a low cost. The sequencing of large amounts of genomic and proteomic data empowers scientific achievements, many of which were thought to be impossible, and novel biological applications have been developed to study their genetic contribution to human diseases and evolution. This is especially true for uncovering new insights from comparative genomics to the evolution of the disease. For example, NGS allows researchers to identify all changes between sequences in the sample set, which could be used in a clinical setting for things like early cancer detection. This dissertation describes a set of computational bioinformatic approaches that bridge the gap between the large-scale, high-throughput sequencing data that is available, and the lack of computational tools to make predictions for and assist in evolutionary studies. Specifically, I have focused on developing computational methods that enable analysis and visualization for three distinct research tasks. These tasks focus on NGS data and will range in scope from processed genomic data to raw sequencing data, to viral proteomic data. The first task focused on the visualization of two genomes and the changes required to transform from one sequence into the other, which mimics the evolutionary process that has occurred on these organisms. My contribution to this task is DCJVis. DCJVis is a visualization tool based on a linear-time algorithm that computes the distance between two genomes and visualizes the number and type of genomic operations necessary to transform one genome set into another. The second task focused on developing a software application and efficient algorithmic workflow for analyzing and comparing raw sequence reads of two samples without the need of a reference genome. Most sequence analysis pipelines start with aligning to a known reference. However, this is not an ideal approach as reference genomes are not available for all organisms and alignment inaccuracies can lead to biased results. I developed a reference-free sequence analysis computational tool, NoRef, using k-length substring (k-mer) analysis. I also proposed an efficient k-mer sorting algorithm that decreases execution time by 3-folds compared to traditional sorting methods. Finally, the NoRef workflow outputs the results in the raw sequence read format based on user-selected filters, that can be directly used for downstream analysis. The third task is focused on viral proteomic data analysis and answers the following questions: 1. How many viral genes originate as "stolen host" (human) genes? 2. What viruses most often steal genes from a host (human) and are specific to certain locations within the host? 3. Can we understand the function of the host (human) gene through a viral perspective? To address these questions, I took a computational approach starting with string sequence comparisons and localization prediction using machine learning models to create a comprehensive community data resource that will enable researchers to gain insights into viruses that affect human immunity and diseases.

Probabilistic models are becoming increasingly important in analysing the huge amount of data being produced by large-scale DNA-sequencing efforts such as the Human Genome Project. For example, hidden Markov models are used for analysing biological sequences, linguistic-grammar-based probabilistic models for identifying RNA secondary structure, and probabilistic evolutionary models for inferring phylogenies of sequences from different organisms. This book gives a unified, up-to-date and self-contained account, with a Bayesian slant, of such methods, and more generally to probabilistic methods of sequence analysis. Written by an interdisciplinary team of authors, it aims to be accessible to molecular biologists, computer scientists, and mathematicians with no formal knowledge of the other fields, and at the same time present the state-of-the-art in this new and highly important field.

In recent decades, new technologies have made remarkable progress in helping to understand biological systems. Rapid advances in genomic profiling techniques such as microarrays or high-performance sequencing have brought new opportunities and challenges in the fields of computational biology and bioinformatics. Such genetic sequencing techniques allow large amounts of data to be produced, whose analysis and cross-integration could provide a complete view of organisms. As a result, it is necessary to develop new techniques and algorithms that carry out an analysis of these data with reliability and efficiency. This Special Issue collected the latest advances in the field of computational methods for the analysis of gene expression data, and, in particular, the modeling of biological processes. Here we present eleven works selected to be published in this Special Issue due to their interest, quality, and originality.

This book contains the latest material in the subject, covering next generation sequencing (NGS) applications and meeting the requirements of a complete semester course. This book digs deep into analysis, providing both concept and practice to satisfy the exact need of researchers seeking to understand and use NGS data reprocessing, genome assembly, variant discovery, gene profiling, epigenetics, and metagenomics. The book does not introduce the analysis pipelines in a black box, but with detailed analysis steps to provide readers with the scientific and technical backgrounds required to enable them to conduct analysis with confidence and understanding. The book is primarily designed as a companion for researchers and graduate students using sequencing data analysis but will also serve as a textbook for teachers and students in biology and bioscience.

The State of the Art in Transcriptome AnalysisRNA sequencing (RNA-seq) data offers unprecedented information about the transcriptome, but harnessing this information with bioinformatics tools is typically a bottleneck. RNA-seq Data Analysis: A Practical Approach enables researchers to examine differential expression at gene, exon, and transcript le

This textbook provides step-by-step protocols and detailed explanations for RNA Sequencing, ChIP-Sequencing and Epigenetic Sequencing applications. The reader learns how to perform Next Generation Sequencing data analysis, how to interpret and visualize the data, and acquires knowledge on the statistical background of the used software tools. Written for biomedical scientists and medical students, this textbook enables the end user to perform and comprehend various Next Generation Sequencing applications and their analytics without prior understanding in bioinformatics or computer sciences.