DNA and RNA isolation techniques for non-experts / Akash Gautam.

Format:

Book

Author/Creator:

Gautam, Akash, author.

Series:

Techniques in Life Science and Biomedicine for the Non-Expert

Language:

English

Subjects (All):

Nucleotide sequence.

Physical Description:

1 online resource (203 pages)

Place of Publication:

Cham, Switzerland : Springer, [2022]

Summary:

This thorough introductory volume presents the background, applications, and stepwise directions for standard DNA and RNA isolation techniques. Unlike a kit chemistry approach, this book provides a breadth of information necessary for junior or non-expert researchers to learn and apply these techniques in their work. An accessible, indispensable how-to guide for researchers in immunology, molecular biology, zoology, forensic science, genetics, botany, neuroscience, physiology, and others.

Contents:

Intro

Sources and Acknowledgments

Contents

Contributors

Part I: Introductory Chapters

Basic Idea About Nucleic Acids for Nonexperts

1 Molecules of Life

2 Nucleic Acids

3 DNA

3.1 Secondary Structure of DNA: The Double Helix

3.2 Other Secondary Structures of DNA/Types of DNA

3.3 Physical and Chemical Properties of DNA

3.4 From DNA to Chromosome

3.5 Functions of DNA

4 RNA

4.1 Structure

4.2 Types of RNA

4.3 Functions of RNA

References

General Steps During Isolation of DNA and RNA

1 Introduction

2 Common Steps of Nucleic Acid Isolation

3 Widely Used Extraction Methods

4 DNA Extraction and Purification Based on Source Materials

4.1 DNA Isolation from Microbes

4.2 DNA Extraction from Animal Cells and Tissues

4.3 DNA Extraction from Plant Tissues and Cells

5 RNA Isolation

Part II: Methods

Phenol-Chloroform DNA Isolation Method

1 Introduction of the Technique

2 Basic Principle

3 Protocol

3.1 Reagents Required

3.2 Working Solutions

3.3 Procedure

3.3.1 Sample Collection

3.3.2 Protocol for DNA Isolation from Whole Blood (20 mL)

4 Precautions

5 Applications

5.1 Gene Sequencing

5.2 Genetic Engineering

RNA Isolation by the Guanidinium-Acid-Phenol Method

Spin Column-Based Isolation of Nucleic Acid

3.2 Working Solution

Isolation of Plasmid DNA by Alkaline Lysis

3.3 Procedure.

3.3.1 Agarose Gel Electrophoresis

3.3.2 Determination of Plasmid Concentration by Spectrophotometry

DNA Isolation by Hydrophilic Ionic Liquid Treatment

3.3.1 Procedure of DNA Extraction

5.1 Food Safety Surveillance

5.2 PCR Enhancement

Lithium Chloride-Based Isolation of RNA

Isolation of Bacteriophage DNA by PEG Method

3.3.1 Precipitation of Bacteriophage Particle with PEG

3.3.2 Extraction of Phage DNA with Phenol

3.3.3 Precipitation of Phage DNA

5.1 Genetic Engineering

5.2 Molecular Studies

5.3 Vaccine Development

DNA Isolation by Chelex Method

1.1 Basic Principle

2 Protocol

2.1 Reagents Required

2.2 Working Solutions

2.3 Procedure

2.3.1 Preparation of Blood and Tissue Sample

2.3.2 DNA Isolation Protocol

2.3.3 Protein Precipitation

2.3.4 DNA Precipitation

3 Precautions

4 Applications

4.1 Forensic Science

4.2 Pathology

4.3 Gene Sequencing

DNA Isolation by Lysozyme and Proteinase K

3.3 Protocol for DNA Isolation

Isolation of DNA from Blood Samples by Salting Method

3 Protocol.

3.1 Reagents Required

3.3 Procedure for DNA Extraction

3.3.1 RBC Lysis

3.3.2 WBC Lysis

3.3.3 Precipitation of DNA

3.3.4 Washing of DNA

3.3.5 Storage of DNA

CTAB or SDS-Based Isolation of Plant's DNA

3.3.1 The Other Two Protocols with Some Variations Are Also Discussed Below

DNA Extraction by Spooling Method

Magnetic Bead-Based Nucleic Acid Isolation

Density Gradient-Based Nucleic Acid Isolation

3.3 Protocol

DNA Extraction from Agarose Gel Through Paper Strips

3.1 Reagent Required

3.2.1 Preparation of 2% Agarose Gel

3.2.2 Restriction Digestion Reaction

Part III: Major Applications of DNA and RNA

Transformation or Genetic Modification of Cells/Organisms

2 Natural Method of Genetic Modification

3 Genetic Modification in Plants

4 Genetic Modification in Animals and Microbes

5 Non-targeted Mutagenesis

6 Genetic Engineering

Gene Cloning and Vectors.

1 Introduction

2 Basic Steps of the Technique (Cell-Based DNA Cloning)

3 Vector or Cloning Vector

4 Types of Cloning Vectors

Polymerase Chain Reaction (PCR)

1 Concept of PCR

2 Taq Polymerase

3 Primers

4 Buffer System

5 The Reaction Conditions

6 Basic Principle

7 Short Protocol

8 Applications

Southern and Northern Blotting

2 Materials Required

3 General Protocol

3.1 Restriction Digestion and Gel Electrophoresis

3.2 Transfer

3.3 Prehybridization

3.4 Hybridization

3.5 Detection of Probe

4 Application

Genome Mapping

2 Difference Between Genome Mapping and DNA Sequencing

3 Types of Genome Mapping

4 Genetic Mapping

5 Physical Mapping

Applications of DNA Sequencing Technologies for Current Research

2 History

3 Basic Principle

4 Maxam-Gilbert Sequencing

5 Chain-Termination or Dideoxy Method/Sanger Sequencing Principle

6 Large-Scale Sequencing and De Novo Sequencing

7 Shotgun Sequencing

8 Clone Contig Approach

9 Amplicon Sequencing

10 High-Throughput Methods

11 Long-Read Sequencing Methods: Nanopore Sequencing

12 Short-Read Sequencing Methods: 454 Pyrosequencing

13 Illumina (Solexa) Sequencing

14 SOLiD Sequencing

15 Ion Torrent Semiconductor Sequencing

16 PacBio Sequencing

17 The Basic Protocol for DNA Sequencing

18 Precautions

19 Need of DNA Sequencing Technology

20 Conclusion

Index.

Notes:

Includes bibliographical references and index.

Description based on print version record.

Other Format:

Print version: Gautam, Akash DNA and RNA Isolation Techniques for Non-Experts

ISBN:

9783030942304