Syllabus
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Course Code: IN-BTY-501 Course Name: Recombinant DNA Technology |
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| MODULE NO / UNIT | COURSE SYLLABUS CONTENTS OF MODULE | NOTES |
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| 1 | Genetic Engineering: Introduction, scope and applications of Genetic Engineering, Miles stones in Genetic engineering, Central role of E.coli. Gene Modifying Enzymes: DNA polymerase, Polynucleotide kinase, Terminal deoxynucleotidy1 transferase, Reverse transcriptase, Restriction endonucleases (R.E.) - Host controlled restriction and modification, Nomenclature, types, Recognition sequence, blunt and sticky ends, applications, online tools & Database (like REBASE) for studyingrecognition site and about R.E. enzymes, Nucleases, Methylases, Alkaline phosphatase, Ligases- E. coli and T4 DNA ligases, Linker, Adaptor, Homopolymer tailing, Nick translation system Gene Cloning Vectors: Types, classes and uses of cloning vectors viz. Plasmids, Cosmids, Phagemid Plasmid Biology: Structural and Functional Organization of Plasmids, Plasmid Replication, Stringent and Relaxed Plasmids, Incompatibility of Plasmid Maintenance, Ti plasmids, Biology of Bacteriophage Lambda: Lambda Phage as a natural in vivo vector, in vitro construction of lambda vector, Bacteriophage (ssDNA Phages), Cauliflower Mosaic Virus, Artificial chromosomes (YAC, BAC, PAC). |
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| 2 | Isolation & Analysis of Genomic and Nuclear DNA: Purification of total cell DNA, plasmid DNA, phage DNA, DNA digestion and restriction fragment analysis and sequencing by chemical, enzymatic and BigDye terminator methods. Polymerase Chain Reaction: Concept, Basic PCR reaction, Factors affecting the PCR, Types of PCR (RT- PCR, Real time PCR, Allele specific PCR, Multiplex PCR), Applications of PCR, Preparation of Primers by using online tools Cloning and Subcloning Strategy: Construction of recombinant DNA: Preparation of competent cell - Transformation, Transfection – Selection and Screening of Recombinants (bacteria and phages); Cloning strategies in yeast, E. coli and B. subtilis |
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