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BIO 603 GENETIC ENGINEERING – PRINCIPLES AND METHODS                                                                               (3L +1P) I

Objective
  To provide knowledge about the basics of recombinant DNA technology and its applications

Theory

UNIT I
  Introduction and historical perspectives; Properties and applications of Restriction Enzymes; Characteristics of vectors; DNA cloning strategies.

UNIT II
  Methods of gene isolation, Construction and screening of genomic and cDNA libraries, PCR; Site directed mutagenesis; Gene silencing Methodologies:–Anti-sense RNA Technology, Ribozyme technology; DNA sequencing.

UNIT III
  Plant transformation methods and transgene analysis, Potential applications of genetic engineering in agriculture; Transcriptional and post transcriptional gene silencing, gene targeting and gene therapy.

UNIT IV
  Bioethics and biosafety issues and IPR in Recombinant DNA research.

Practicals
  Isolation of Plant, plasmid and phage DNA and purification; Restriction of Plant DNA; Southern blotting; Elution of DNA from agarose gel; Radioactive labeling of DNA; Southern hybridization, autoradiography. Preparation of vector for cloning; Ligation of vector with insert DNA; Preparation of competent cells; Transformation of E.coli cells and selection of recombinants by antibiotic or blue/white selection; PCR, Preparation of plant RNA, RT-PCR.

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