Course

MOLECULAR TOOLS FOR GENE CLONING

Nucleases: Exonucleases and Endonucleases, Restriction Enzymes (Type I, Type II, Type III, Type IV & Type V), RNases
Methylases: CpG Methylase , Dam Methylase, Dcm Methylase
Polymerases: DNA Pol I, Klenow Fragments, Reverse Transcriptase, Taq & Pfu Polymerases
Ligases: T4 DNA Ligase, E.coli DNA Ligase, T4 RNA Ligase
Topoisomerases: Type I(A, B) & Type II(A,B)
End Modifying Enzymes: Terminal Transferase, T4 Polynucleotide Kinase, Alkaline Phosphatases

VECTORS AND GENE CLONING

Introduction to cloning vectors — Desirable properties of vectors – Prokaryotic & Eukaryotic Expression Systems (Constitutive & Inducible)
Plasmid Vectors — Phage Vectors — Cosmids — Phagemids — BACs — Yeast Vectors — YACs — Lentiviral Vectors — Adenoviral Vectors — Plant Vectors
Insect Vectors (Optional Reading)

ADVANCED TECHNIQUES IN MOLECULAR BIOLOGY

Polymerase Chain Reaction — Quantitative Real Time PCR — Gel Electrophoresis: AGE & PAGE — Blotting Techniques: Southern, Western & Northern
Methods of gene transfer in Plants and Animals: Chemical, Physical & Viral mediated DNA transfer
Construction of Genomic & cDNA Libraries — DNA Sequencing — Protein Engineering: Site Directed Mutagenesis — Reporter Gene Assays — DNA Protein Interactions: EMSA, DNA Footprinting — Protein Protein Interactions: Y2H, Y3H, B1H, B2H

RECENT TRENDS IN MOLECULAR BIOLOGY RESEARCH

Targeted Genome Editing: ZFNs, TALENs, CRISPRs — Gene Targeting: Knock-ins & Knock-outs — DNA Finger Printing

1. Principles of Gene Manipulation and Genomics – 7^th Edition – Sandy B. Primrose, Richard Twyman – Blackwell Publishing
2. Gene Cloning and DNA Analysis: An Introduction – 6th Edition – T. A. Brown – John Wiley & Sons
3. An Introduction to Genetic Engineering – 3rd Edition – Desmond S. T. Nicholl – Cambridge University Press
4. Molecular Biotechnology: Principles and Applications of Recombinant DNA – 4th Edition – Bernard R. Glick, Jack J. Pasternak, Cheryl L. Patten – ASM Press

• A PBS Documentary entitled “Playing God” [History of Genetic Engineering]
• What is Genetic Engineering? [A short intro to the subject]