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Module I:Introduction to Cloning(4hrs)
Overview of Cloning, Purification and Separation of Nucleic Acids – cutting and joining DNA and vectors. Plasmid vectors, phage vectors,cosmids .
Practice: Restriction enzyme digestion of plasmid DNA or lambda DNA
Module II: Genetic manipulation(4hrs)
Manipulation of DNA–Restriction and Modification enzymes, Design of linkers and adaptors.
Practice:Gel purification of RE digested DNA
Module III: Cloning Vectors and their application(4hrs)
Characteristics of cloning and expression vectors based on plasmid and bacteriophage, Vectors for yeast, insect and mammalian systems, Prokaryotic and eukaryotic expression host systems
Practice: Ligation of DNA fragments with cloning vector pUC18 or pBR322.
Module IV: DNA sequencing(5hrs)
Maxxam-Gilbert- Sanger methods. Automated DNA sequencing. PCR technology - concept- types- primer design- analysis of products and applications. DNA-finger printing, SYBR green assay, Taqman assay, Molecular beacons.
Practice:Preparation of competent cells and Transformation into E.coli with recombinant vector.
Module V: cDNA arrays and Micro array technology(4 hrs)
Strategies for the production of recombinant proteins - insulin- human growth hormone- industrially important proteins. Construction of genomic library- cDNA library
Practice:Isolation of recombinants and confirmation of insert DNA in vector.
Module VI:Analysis and manipulation of Gene expression(4hrs)
Analysis of gene expression, analyzing transcription and translation, Analysis of gene function, site Directed Mutagenesis,Transposon Mutagenesis
Practice:Primer design for PCR and amplification of DNA by PCR.
Module VII: Cloning Application(5hrs)
applications – vaccines – human and genetic diseases – transgenics, Small double stranded RNAs; siRNA technology; Micro RNA; Construction of siRNA vectors; Principle and application of gene silencing.
Analyzing DNA and cloning
The principles of genetic engineering
Different types of cloning
Impact of cloning on medicine
Studies on DNA-modified organisms
Gene-editing technologies, explaining their impact on genetic engineering
Cloning: positive or negative outcome for future generations?
Human Genome Project.