Introduction to Unit 13 : Important Topics To Read
Unit 13: Last but not least! Dealing with all different techniques and their applications, extremely experimental questions are expected from this unit, which will test your true research abilities. Scoring cannot be guaranteed from 1st 3 subunits, but you do have some direct questions being asked from the next four subunits. Being very vast, you might face some serious preparation issues, but preparing the following may give you an edge.
SYLLABUS
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 of gene expression at RNA and protein level, large scale expression, 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, fluocytometry and immunofluorescence microscopy, detection of molecules in living cells, in situ localization by techniques such as FISH and GISH.
C Biophysical Method:
- Molecular analysis using UV/visible, fluorescence, circular dichroism, NMR and ESR spectroscopy Molecular structure determination using X-ray diffraction and NMR, Molecular analysis using light scattering, different types of mass spectrometry and surface plasma resonance methods.
D Statisitcal 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. Radiolabeling techniques:
- Detection and measurement of different types of radioisotopes normally used in biology, incorporation of radioisotopes in biological tissues and cells, molecular imaging of radioactive material, safety guidelines.
F. Microscopic techniques:
- Visulization 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 freezefracture 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.
Important Topics to be studied From CSIR NET Unit 13
- Recombinant DNA technology: RNA, DNA and proteins isolation, separation and analytical techniques (1D and 2D gel electrophoresis, isoelectric focusing gels), cloning,
- Recombinant protein expression using various bacterial, animal and plant vectors (cosmid, BAC and YAC vectors), genomic and cDNA libraries,
- In vitro mutagenesis and deletion techniques, gene knock out, DNA and protein sequencing, gene expression analysis at RNA and protein level, microarrays, RFLP, RAPD, and AFLP techniques
- Immunological techniques such as ELISA, RIA, western blot, immunoprecipitation, flow cytometry, immunofluorescence microscopy, FISH and GISH.
- Biophysical methods such as UV/visible, fluorescence spectroscopy, circular dichroism, NMR, ESR spectroscopy, X-ray diffraction, NMR, different types of mass spectrometry.
- Statistical Methods – Mainly questions will be asked from probability distributions such as Binomial, Poisson and normal.
- T-test, X2 test
- Radiolabeling techniques – Various types of radioisotopes used in biology and their properties focus on their detection, measurement, and incorporation
- Microscopic techniques – numerical on resolving powers of different microscopes, SEM, TEM, fixation, and staining techniques, freeze-etch and freeze-fracture methods for EM
- Electrophysiological methods – Single neuron recording, patch-clamp recording, ECG, Brain activity recording, PET, MRI, fMRI, CAT
REFERENCE BOOKS For CSIR NET UNIT 13
- Principles of Gene Manipulation and Genomics by Sandy B. Primrose, Richard Twyman
- Gene Cloning and DNA Analysis: An Introduction by T. A. Brown
- Molecular Cloning: A Laboratory Manual by Joseph Sambrook, David Russell
- Fundamentals of Biostatistics by V.B. Rastogi
- Principles and Techniques of Biochemistry and Molecular Biology by Wilson/Walker
