Argonaute Proteins: Methods and Protocols by Katsutomo Okamura (English) Hardcov

Publisher Description

This volume covers methods that analyze various Argonaute proteins from a variety of organisms to help researchers better understand their properties ranging from a molecular level to an organismal level. The chapters in this book explore the following topics: identification and expression analysis of guide nucleic acids and their targets; analysis of biochemical properties of Argonautes; biological functions of Argonautes; and obtaining materials and setting up analysis platforms. 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. Cutting-edge and authoritative, Argonaute Proteins: Methods and Protocols is a valuable resource for researchers and scientists looking to expand their knowledge ofArgonaute proteins and their functions.

Back Cover

This volume covers methods that analyze various Argonaute proteins from a variety of organisms to help researchers better understand their properties ranging from a molecular level to an organismal level. The chapters in this book explore the following topics: identification and expression analysis of guide nucleic acids and their targets; analysis of biochemical properties of Argonautes; biological functions of Argonautes; and obtaining materials and setting up analysis platforms. 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. Cutting-edge and authoritative, Argonaute Proteins: Methods and Protocols is a valuable resource for researchers and scientists looking to expand their knowledge of Argonaute proteins and their functions.

Table of Contents

Cloning and Identification of Recombinant-Argonaute Bound Small RNAs using Next-Generation Sequencing.- Quantification of miRNAs Co-Immunoprecipitated with Argonaute Proteins using SYBR Green-Based qRT-PCR.- Gateway to Understanding Argonaute Loading of Single-Stranded RNAs: Preparation of Deep Sequencing Libraries with In Vitro Loading Samples.- Dumbbell-PCR for Discriminative Quantification of a Small RNA Variant.- MicroRNA Detection by Whole-Mount In Situ Hybridization in C. elegans.- cCLIP-Seq: Retrieval of Chimaeric Reads from HITS-CLIP (CLIP-Seq) Libraries.- Kinetic Analysis of Small Silencing RNA Production by Human and Drosophila Dicer Enzymes In Vitro.- Nucleic Acid-Binding Assay of Argonaute Protein using Fluorescence Polarization.- Reconstitution of RNA Interference Machinery.- Single-Molecule Analysis for RISC Assembly and Target Cleavage.- Profiling Open Chromatin Structure in the Ovarian Somatic Cells using ATAC-Seq.- Assessing miR-451 Activity and Its Role in Erythropoiesis.- Functional Analysis of MicroRNAs in Neurogenesis during Mouse Cortical Development.- Cellular Approaches in Investigating Argonaute2-Dependent RNA Silencing.- Genomic Tagging of AG01 using CRISPR/Cas9-Mediated Homologous Recombination.- Accurate Profiling and Quantification of tRNA Fragments from RNA-Seq Data: A vade mecum for MINTmap.

Feature

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

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