Unit 13: Methods in biology
A. Molecular biology and recombinant DNA methods: Isolation and purification of RNA, DNA (genomic and plasmid) and proteins, different separation methods; analysis of RNA, DNA and proteins by one and two dimensional gel electrophoresis, isoelectric focusing gels; molecular cloning of DNA or RNA fragments in bacterial and eukaryotic systems; expression of recombinant proteins using bacterial, animal and plant vectors; isolation of specific nucleic acid sequences; generation of genomic and cDNA libraries in plasmid, phage, cosmid, BAC and YAC vectors; in vitro mutagenesis and deletion techniques, gene knock out in bacterial and eukaryotic organisms; protein sequencing methods, detection of post-translation modification of proteins; DNA sequencing methods, strategies for genome sequencing; methods for analysis, such as micro array based techniques; isolation, separation and analysis of carbohydrate and lipid molecules; RFLP, RAPD and AFLP techniques.
B. Histochemical and immunotechniques: Antibody generation, detection of molecules using ELISA, RIA, western blot, immunoprecipitation, floweytometry and immunofluorescence microscopy, detection of molecules in living cells, in situ localization by techniques such as FISH and GISH.
C. Biophysical Methods: Analysis of biomolecules using UV/Visible, fluorescence, circular dicroism, NMR and ESR spectroscopy, structure determination using X-ray diffraction and NMR; analysis using light scattering, different types of mass spectrometry and surface plasma resonance methods.
D. Statistical Methods: Measures of central tendency and dispersal; probability distributions Binomial, Poisson and normal); sampling distribution; difference between parametric and non-parametric statistics; confidence interval; errors; levels of significance; regression and correlation; t-test; analysis of variance; X² test; basic introduction to Muetrovariate statistics, etc.
E. Radio-labeling Techniques: Properties of different types of radioisotopes normally used in biology, their detection and measurement; incorporation of radioisotopes in biological tissues and cells, molecular imaging of radioactive material, safety guidelines.
F. Microscopic techniques: Visualization of cells and subcellular components by light microscopy, resolving powers of different microscopes, microscopy of living cells, scanning and transmission microscopes, different fixation and staining techniques for EM, Freeze-etch and freeze-fracture methods for EM, image processing methods in microscopy.
G. Electrophysiological methods: Single neuron recording, patch-clamp recording, ECG, Brain activity recording, lesion and stimulation of brain, pharmacological testing, PET, MRI, fMRI, CAT.
H. Methods in field biology: Methods of estimating population density of animals and plants, ranging patterns through direct, indirect and remote observations, sampling methods in the study of behavior, habitat characterization-ground and remote sensing methods.
I. Computational Methods: Nucleic acid and protein sequence databases; data mining methods for sequence analysis, web-based tools for sequence searches, motif analysis and presentation.
A. Molecular biology and recombinant DNA methods: Isolation and purification of RNA, DNA (genomic and plasmid) and proteins, different separation methods; analysis of RNA, DNA and proteins by one and two dimensional gel electrophoresis, isoelectric focusing gels; molecular cloning of DNA or RNA fragments in bacterial and eukaryotic systems; expression of recombinant proteins using bacterial, animal and plant vectors; isolation of specific nucleic acid sequences; generation of genomic and cDNA libraries in plasmid, phage, cosmid, BAC and YAC vectors; in vitro mutagenesis and deletion techniques, gene knock out in bacterial and eukaryotic organisms; protein sequencing methods, detection of post-translation modification of proteins; DNA sequencing methods, strategies for genome sequencing; methods for analysis, such as micro array based techniques; isolation, separation and analysis of carbohydrate and lipid molecules; RFLP, RAPD and AFLP techniques.
B. Histochemical and immunotechniques: Antibody generation, detection of molecules using ELISA, RIA, western blot, immunoprecipitation, floweytometry and immunofluorescence microscopy, detection of molecules in living cells, in situ localization by techniques such as FISH and GISH.
C. Biophysical Methods: Analysis of biomolecules using UV/Visible, fluorescence, circular dicroism, NMR and ESR spectroscopy, structure determination using X-ray diffraction and NMR; analysis using light scattering, different types of mass spectrometry and surface plasma resonance methods.
D. Statistical Methods: Measures of central tendency and dispersal; probability distributions Binomial, Poisson and normal); sampling distribution; difference between parametric and non-parametric statistics; confidence interval; errors; levels of significance; regression and correlation; t-test; analysis of variance; X² test; basic introduction to Muetrovariate statistics, etc.
E. Radio-labeling Techniques: Properties of different types of radioisotopes normally used in biology, their detection and measurement; incorporation of radioisotopes in biological tissues and cells, molecular imaging of radioactive material, safety guidelines.
F. Microscopic techniques: Visualization of cells and subcellular components by light microscopy, resolving powers of different microscopes, microscopy of living cells, scanning and transmission microscopes, different fixation and staining techniques for EM, Freeze-etch and freeze-fracture methods for EM, image processing methods in microscopy.
G. Electrophysiological methods: Single neuron recording, patch-clamp recording, ECG, Brain activity recording, lesion and stimulation of brain, pharmacological testing, PET, MRI, fMRI, CAT.
H. Methods in field biology: Methods of estimating population density of animals and plants, ranging patterns through direct, indirect and remote observations, sampling methods in the study of behavior, habitat characterization-ground and remote sensing methods.
I. Computational Methods: Nucleic acid and protein sequence databases; data mining methods for sequence analysis, web-based tools for sequence searches, motif analysis and presentation.
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