Contents

1. Introduction to Recombinant DNA Technology

1. Historical Perspective

2. Concept of Recombinant DNA Technology

2.1 Significance of Recombinant DNA Technology

3. Basic Layout of r-DNA Laboratory

3.1 Layout of Recombinant DNA Laboratory 

2. Molecular Tools used in Recombinant DNA Technology

1. DNA Modifying Enzymes

2. Restriction Enzymes

2.1 Types of Restriction Enzymes

2.2 Nomenclature of Restriction Enzymes

2.3 Restriction Enzyme Recognition Sequences

2.4 Properties of Restriction Enzymes

3. DNA Ligases

4. DNA Polymerases

4.1 DNA Dependent DNA Polymerases I

5. Alkaline Phosphatase

6. Nucleases-Mode of Action and Applications 

3. Vectors used in Recombinant DNA Technology

1. Introduction

2. Plasmid Vectors

2.1 Properties of a Good Plasmid

2.2 Nomenclature of Plasmid Vectors

2.3 Types of Bacterial Plasmids

3. Phage

3.1 Structure of a Typical Phage

3.2 Lambda (l)

3.3 Why Bacteriophage Lambda is considered a Good Cloning Vehicle?

3.4 Phage M13

4. Expression Vector

5. Agrobacterium based Plasmid Vectors

6. Chimeric Vectors, Cosmids, Phasmids Phagemids

6.1 Cosmid Vectors

6.2 Phasmids

6.3  Phagemids

7. Yeast Artificial Chromosome (YAC) Vectors

8. Bacterial Artificial Chromosome (BAC) Vectors 

4. Construction of Genomic and cDNA Library

1. Introduction

2. Genomic Library

2.1 Screening of Recombinants

3. The cDNA Library

3.1 THE cDNA Library Construction

3.2 Cloning of cDNA

4. Comparison between cDNA Library and Genomic Library

5. Applications of Genomic and cDNA Libraries 

5. Polymerase Chain Reaction (PCR)

1. Introduction

2. Steps involved in PCR

3. Applications of PCR

4. Real Time PCR 

6. Sequencing of Genes and Genomes

1. Introduction

2. Maxam and Gilbert’s Chemical Degradation Method

3. Sanger’s Enzymatic Method (Sanger’s Dideoxy Method)

4. Automated DNA Sequencing

5. Next Generation Sequencing 

7. Applications of Recombinant DNA Technology

1. Introduction

2. Human Insulin

3. Gene Therapy

3.1 Human Growth Hormone (HGH)

3.2 Bovine Somatotrophin (BST)

3.3 Rennin

4. Introduction to CRISPR/Cas9 as Genome Editing Tool