http://www.loc.gov/mods/v3 http://www.loc.gov/standards/mods/v3/mods-3-2.xsd
High-density sequencing applications in microbial molecular genetics
High-Density Sequencing Applications in Microbial Molecular Genetics, Volume 612 in the Methods of Enzymology series provides the latest on the high-density sequencing of DNA and cDNA libraries and how they have revolutionized contemporary research in biology. Methods permitting tens of millions of sequence reads in a single experiment have paved the way to genome-wide studies that are contributing to our understanding of the complexity of living systems. Chapters in this updated volume include Characterizing the role of exoribonucleases in the control of microbial gene expression: Differential RNA seq., Conformational studies of bacterial chromosomes by high-throughput sequencing methods, Measuring mRNA degradation, and more. Addition sections cover Global recognition patterns of bacterial RNA-binding proteins, High-resolution profiling of NMD targets, and the Generation of a metagenomic 3C/Hi-C library of human gut microbiota, Genome-wide mapping of yeast retrotransposons integration target sites, Measuring protein synthesis rates, Finding unsuspected partners of small RNAs with new screening approaches, Use of multiplexed transcriptomics to define the relationship between promoter sequence and transcription output, RNA-based control of quorum sensing in Vibrio cholerae, amongst other highly regarded topics. Detail methods used in research articles that were recently published in leading journals Provides the latest on the high-density sequencing of DNA and cDNA libraries and how they have revolutionized contemporary research in biology.
Front Cover; High-Density Sequencing Applications in Microbial Molecular Genetics; Copyright; Contents; Contributors; Preface; Chapter One: Characterizing the Role of Exoribonucleases in the Control of Microbial Gene Expression: Differential RNA-Seq; 1. Overview of Exoribonucleases; 2. Overview of Differential RNA-Seq; 3. RNA Extraction; 4. Bioinformatics Analysis; 4.1. Mapping of Reads; 4.2. Transcript Expression Quantification; 4.3. Differential Expression Analysis; 4.4. Functional Annotation Analysis; 5. Experimental Validation; 5.1. qPCR; 5.2. Motility Assays; 6. Conclusion
AcknowledgmentsReferences; Chapter Two: Conformational Studies of Bacterial Chromosomes by High-Throughput Sequencing Methods; 1. Introduction; 1.1. Chromosome Conformation; 1.2. DNA-Protein Interactions; 1.3. Combining NGS to 3C and ChIP Approaches; 2. 3C-Seq: The Protocol; 2.1. Buffers and Reagents; 2.2. Cross-Linking of Cell Cultures; 2.3. 3C Library Preparation; 2.3.1. Cell Lysis; 2.3.2. RE Digestion; 2.3.3. Ligation; 2.3.4. DNA Purification; 2.4. Notes; 3. ChIP-Seq: The Protocol; 3.1. Buffers and Reagents; 3.2. Cross-Linking of Cell Cultures; 3.3. Cell Lysis
3.4. Batch Immunoprecipitation of FLAG Fusions Proteins3.5. Elution of the Immunoprecipitated Protein; 3.6. Purification of the Coimmunoprecipitated DNA; 3.7. Notes; 4. Data Processing and Analysis; 4.1. Analysis of 3C-Seq Data; 4.2. Analysis of ChIP-Seq Data; 5. Conclusion; Acknowledgments; References; Chapter Three: Large-Scale Measurement of mRNA Degradation in Escherichia coli: To Delay or Not to Delay; 1. Introduction; 2. Measuring mRNA Degradation Rates; 2.1. Methods; 2.2. Rifampicin; 2.3. Practical Considerations; 3. Genome-Wide mRNA Quantification
3.1. mRNA Extraction and Quantification3.2. Normalization; 4. mRNA Half-Life Estimation; 4.1. Decay Models; 4.2. Accuracy Criteria of the Estimation; 5. Effect of the Delay Before the Onset of Exponential Decay on mRNA Half-Life Estimation; 5.1. Datasets of mRNA Levels Over the Time; 5.2. Estimations of the Delay Before the Onset of the mRNA Exponential Decay; 5.3. Comparison of mRNA Half-Lives Estimated With and Without a Delay; 5.4. Correlation Between mRNA Half-Lives Estimated With and Without a Delay; 5.5. Influence of the Delay Extent
5.6. Validation on RT-qPCR mRNA Quantification Datasets6. Concluding Remarks; Acknowledgments; References; Chapter Four: FASTBAC-Seq: Functional AnalysiS of Toxin-Antitoxin Systems in BACteria by Deep Sequencing; 1. Introduction; 2. Materials; 2.1. Molecular Biology Equipment; 2.2. General Equipment for Bacterial Cultures; 2.3. E. coli-Specific Equipment; 2.4. H. pylori-Specific Equipment; 2.5. Molecular Biology Reagents; 2.6. E. coli Strains and Cell Culture; 2.7. H. pylori Strains and Cell Culture
Cambridge, MA : Academic Press, an imprint of Elsevier, 2018