MB 504 TECHNIQUES IN MICROBIOLOGY                                                                                 (1L +2P) I

To familiarize the students with basic safety rules in the microbiological laboratory and to introduce the concepts involved in different methods and techniques applied in microbiological sciences


An introduction to laboratory instruments, Safety rules in laboratory, Handling of  different glassware, Handling of autoclave, Hot air oven, BOD incubator, Laminar flow, Colony counter, pH meter, Biological filters, Spectrophotometer, Microscopy: Light, Compound, Darkfield, Phase Contrast, Fluorscent, EM, TEM, SEM.

Culture: Types of culture and culture media, Inoculation techniques, Isolation of pure culture, Proof of purity of cultures, Maintenance and preservation of pure cultures, Culture collection: World scenario and Indian scenario, Techniques for microscopic examination of  living microorganisms (Bacteria/Cyanobacteria): Wet mount, Hanging drop technique, Stains and staining techniques, Enumeration of bacteria, Cyanobacteria by plate count, Turbidometric method and MPN method.

Identification of bacteria using biochemical tests, Introduction and importance of Bergey’s Manual, Bioassay techniques, Antibiotic sensitivity of bacteria.

Molecular- biology techniques, Isolation of Genomic DNA and PCR amplification in bacteria and cyanobacteria, RAPD and RFLP, Isolation of plasmids, Protein profiling by SDS-PAGE, Transposon and chemical mutagenesis, Biochemical characterization in cyanobacteria.

Introduction to safety measures in laboratory and maintenance of aseptic conditions, Microscopy and instrumentation, Media preparation and sterilization, Isolation and purification techniques, Enumeration and quantification techniques, Staining of micro-organisms: Simple staining, Negative staining, Gram staining, Capsule staining, Spore staining, Motility test, Biochemical characterization of micro-organisms, Identification of unknown bacteria, Carbohydrate fermentation, Triple sugar- iron agar test, IMViC test, Hydrogen sulfide test, Urease test, Litmus milk reaction, Nitrate reduction test, Catalase and Oxidase test, Isolation and purification of cyanobacteria, Identification of cyanobacteria, RAPD analysis in cyanobacteria, Enumeration techniques in cyanobacteria, Estimation of pigments and nitrogen fixation, Antibiotic resistance, Isolation of Genomic DNA and PCR amplification, Restriction digestion of DNA, RFLP analysis, Isolation of plasmids, Protein profiling, Transposon and chemical mutagenesis, Bioinformatic tools.

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