Basic DNA and RNA Protocols 1st edition by Adrian J Harwoo ISBN 089603402X 978-0896034020

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ISBN 10: 089603402X
ISBN 13: 978-0896034020
Author: Adrian J Harwoo 

An essential core collection of the latest molecular and genetic techniques for cloning, subcloning, sequencing, PCR, protein expression, and much more. Each protocol represents a time-tested, step-by-step recipe that creates an understanding of the procedure, easily reproducible results, and confidence that the procedure will work. The collection includes not only many updated and improved classic techniques, but also a powerful group of advanced methods that point to future progress, among them nonisotopic DNA labeling, silver staining, and automatic sequencing. This excellent bench companion will help those who need to learn for the first time how to conduct research on the molecular biology of nucleic acids or those who need to broaden their competence and laboratory skills. Even highly skilled researchers will find many time-saving techniques.

Basic DNA and RNA Protocols 1st Table of contents:

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Chapter 1: Introduction to DNA and RNA

• The Structure and Function of DNA

• The Structure and Function of RNA

• Key Differences Between DNA and RNA

• Overview of Genetic Information Flow: DNA → RNA → Protein

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Chapter 2: DNA Extraction and Purification Protocols

• Principles of DNA Extraction

• Materials and Reagents Required

• Step-by-Step Protocol for DNA Extraction from Animal Cells

• DNA Extraction from Plant Cells

• DNA Extraction from Bacteria

• Quality Control and Assessment of DNA Purity

• Troubleshooting DNA Extraction Issues

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Chapter 3: RNA Extraction and Purification Protocols

• Principles of RNA Extraction

• Materials and Reagents Required

• RNA Isolation from Animal Cells

• RNA Isolation from Plant Cells

• RNA Isolation from Bacteria and Fungi

• Strategies to Prevent RNA Degradation (RNase Inhibition)

• Quality Control of RNA: Integrity and Purity

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Chapter 4: Quantification of DNA and RNA

• UV Spectrophotometry for Quantification

• Fluorometric Methods for DNA/RNA Quantification

• Agarose Gel Electrophoresis for DNA and RNA Visualization

• Calculating Concentration and Purity Ratios (A260/A280)

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Chapter 5: PCR (Polymerase Chain Reaction)

• Principles of PCR

• Materials and Reagents for PCR

• Step-by-Step PCR Protocol

• Optimizing PCR Conditions (Annealing Temperatures, Mg2+ Concentration, etc.)

• Troubleshooting Common PCR Problems

• Quantitative PCR (qPCR) Basics

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Chapter 6: RNA Analysis and Quantification

• Reverse Transcription PCR (RT-PCR)

• Northern Blotting: Protocol and Applications

• RNA Sequencing and Next-Generation Sequencing (NGS) Overview

• RNA Quantification via qRT-PCR

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Chapter 7: DNA Digestion and Cloning Protocols

• Restriction Enzyme Digestion of DNA

• Ligation and Cloning Protocols

• Plasmid Extraction and Purification

• Transformation of Bacteria with Recombinant DNA

• Verification of Cloning via PCR and Sequencing

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Chapter 8: Gel Electrophoresis for DNA and RNA Analysis

• Agarose Gel Electrophoresis Protocol for DNA

• Polyacrylamide Gel Electrophoresis (PAGE) for RNA and DNA

• Staining and Visualization Methods (e.g., Ethidium Bromide, SYBR Green)

• Troubleshooting Gel Electrophoresis Issues

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Chapter 9: DNA Sequencing

• Sanger Sequencing Method and Protocol

• High-throughput Sequencing Technologies (NGS)

• Primer Design and Quality Control for Sequencing

• Data Analysis and Bioinformatics Tools for Sequencing

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Chapter 10: RNA Stability and Storage Protocols

• Principles of RNA Stability and Degradation

• Storage Solutions for RNA (Ethanol Precipitation, Drying, etc.)

• Best Practices for Long-Term RNA Storage

• Preventing RNA Degradation in Experimental Settings

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Chapter 11: Applications of DNA and RNA Protocols in Research

• Gene Expression Analysis (e.g., qPCR, Microarray)

• Functional Genomics and RNA Interference (RNAi)

• Genomic Mapping and Sequencing Projects

• CRISPR/Cas9 Gene Editing and RNA Guide Sequences

• Diagnostics: DNA and RNA in Disease Detection

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Chapter 12: Troubleshooting DNA and RNA Protocols

• Common Problems in DNA/RNA Extraction and Purification

• PCR Amplification Failures and Solutions

• Issues with Gel Electrophoresis and Sequencing

• Tips for Improving RNA Yield and Quality

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Appendices

• Glossary of Molecular Biology Terms

• List of Common Reagents and Buffers

• Safety Guidelines for Handling DNA/RNA

• Further Reading and Resources

Index

• Subject Index

• Author Index

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