Animal Biotechnology

M.V.Sc. Programme in Animal Biotechnology as per ICAR guidelines

Minimum Credit Requirements:

S.N.	Subject	Credit Requirement
1	Major	28
2	Minor + Supporting	11
3	Seminar	01
4	Research	20
	TOTAL	60

Major / Minor / Supporting Subjects:

Major Subject	Animal Biotechnology
Minor & Supporting Subject	Veterinary Microbiology, Biochemistry, Animal Reproduction, Animal Genetics and Breeding, Animal Nutrition, Veterinary Physiology

COURSE STRUCTURE

S. N.	Course No.	Title	Credits	Semester	Department (s) offering the course
1	ABT 601	Basic & Applied Biotechnology	3+0=3	I	Animal Biotechnology
2	ABT 602**	Fundamentals of Cell & Molecular Biology	3+0=3	I	Biochemistry
3	ABT 603	Applied Molecular Biology	2+1=3	II	Animal Biotechnology
4	ABT 604**	Animal Cell Culture: Principles and Applications	1+2=3	I	Animal Biotechnology
5	ABT 605**	Molecular Diagnostics	1+2=3	II	Animal Biotechnology
6	ABT 606	Vaccine Biotechnology	2+0=2	III	Animal Biotechnology
7	ABT 607	Immunology Applied to Biotechnology	1+1=2	III	Animal Biotechnology
8	ABT 608	Introduction to Bioinformatics	1+1=2	III	Animal Biotechnology
9	ABT 609**	Animal Genomics	2+1=3	II	Animal Genetics & Breeding
10	ABT 610**	Reproductive Biotechnology	2+1=3	III	Animal Reproduction
11	ABT 611**	Techniques in Molecular Biology and Genetic Engineering	0+3=3	II	Animal Biotechnology
12	ABT 613	Molecular Forensics	2+1=3	III	Animal Biotechnology
13	ABT 616	Animal Biotechnology	3+0=3	II	Animal Biotechnology/ Animal Reproduction/ Animal Genetics & Breeding
14	ABT 691	Master’s Seminar	0+1=1	III
15	ABT 699	Master’s Research	20	III & IV

** Compulsory courses

Semester-wise distribution of courses

S. N.	Course No.	Title	Credits
SEMESTER I
1	ABT 601	Basic & Applied Biotechnology	3+0=3
2	ABT 602**	Fundamentals of Cell & Molecular Biology	3+0=3
3	ABT 604**	Animal Cell Culture: Principles and Applications	1+2=3
SEMESTER II
4	ABT 603	Applied Molecular Biology	2+1=3
5	ABT 605**	Molecular Diagnostics	1+2=3
6	ABT 609**	Animal Genomics	2+1=3
7	ABT 611**	Techniques in Molecular Biology and Genetic Engineering	0+3=3
8	ABT 616	Animal Biotechnology	3+0=3
SEMESTER III
9	ABT 606	Vaccine Biotechnology	2+1=3
10	ABT 607	Immunology Applied to Biotechnology	1+1=2
11	ABT 608	Introduction to Bioinformatics	1+1=2
12	ABT 610**	Reproductive Biotechnology	2+1=3
13	ABT 613	Molecular Forensics	2+1=3
14	ABT 691	Master’s Seminar	0+1=1
SEMESTER IV
15	ABT 699	Master’s Research	20

** The course nos. ABT 602, ABT 604, ABT 605, ABT 609, ABT 610 & ABT 611 (Total credits 18) and Master’s seminar (ABT 691- 0+1) are compulsory. The remaining 10 credits may be offered from any of the available courses as decided by the advisory committee of the student.

Semester-wise distribution of credits

S. N.	Semester	Major		Minor	Total
		Course work	Research
1	I	9	–	6	15
2	II	12	–	3	15
3	III	8	5	2	15
4	IV	–	15	–	15
	TOTAL	29	20	11	60

Basic & Applied Biotechnology

Course No.	ABT-601	Title	Basic & Applied Biotechnology
Credits	3+0=3	Semester	I

Lecture No.	Topic
	UNIT I
1	Historical background of Biotechnology
2 – 3	Scope of Biotechnology
4 – 5	Introduction to Genetic Engineering
6 – 7	Plant tissue culture and its applications
8 – 10	Animal Tissue culture and its applications
11 – 13	Fermentation technology
	UNIT II
14 – 15	Immobilized enzymes
16 – 17	Vaccines – conventional and newer vaccines
18 – 19	Monoclonal antibodies and Hybridoma Technology
20 – 21	Biotechnological tools in development of diagnostic tests
22 – 24	Embryo transfer technology, Sexing of semen, embryos etc
25- 26	Development of transgenic animals and applications
	UNIT III
27 – 28	Study of bacterial/ viral genomes etc
29 – 30	Study of plasmids/phage genome and libraries
31 – 32	Study of physical maps of bacteria,viruses, yeasts etc.
33 – 34	Study of genetic maps, phylogenetic analysis etc.
35 – 36	Different types of DNA markers
37 – 38	Applications of DNA markers in selection of breeds etc.
	UNIT IV
39 – 40	Application of Biotechnology in agriculture
41 – 42	Application of Biotechnology in animal and Veterinary Sciences
43 – 44	Application of Biotechnology in Pharmaceutical Industry
45 – 46	Application of Biotechnology in Food Industry and Chemical Industry
47 – 48	Application of Biotechnology in environmental sciences

Suggested Readings:

Becker, J. M., Cold Well G. A. and Zachgo E. A. 2007. Biotechnology a Laboratory Course, Academic Press

Brown C.M.,Campbell,I.and Priest F. G. 2005. Introduction to Biotechnology. Panima

Singh, B.D. 2006. Biotechnology Expanding Horizon, Kalyani.

Fundamentals of Cell and Molecular Biology

Course No.	ABT 602	Title	Fundamentals of Cello and Molecular Biology
Credits	3 + 0 = 3	Semester	I

THEORY

Lecture Nos.	Topic
	UNIT I
1 – 2	Evolution of cells
3 – 5	Introduction to molecular interactions, thermodynamics and equilibrium in molecular recognition and biological functions.
6 – 8	Energy production: Structure of mitochondria, and chloroplasts, respiratory chain, ATP synthesis, photosynthesis, genomes of mitochondria and chloroplasts,
9 – 12	Cellular compartments and intercellular sorting of proteins: endoplasmic reticulum, lysosome, peroxisomes, synthesis and soorting of proteins lysosomal proteins, membrane proteins, secretary proteins, lipoproteins, glycolipids. Lipid synthesis and transport.
	UNIT II
13 – 16	Cytoskeleton: Mechanism of muscle contraction, actin filaments and cell cortex, cilliary movements and cytoplasmic microtubules and intermediate filaments.
17 – 20	Cell signaling: Endocrine, exocrine and synaptic signaling molecules, surface and intracellular receptors, G proteins and generation of secondary messengers, mode of action of cAMP and Ca++ calmodulin, target cell adaptation.
21 – 24	Cell growth and divisions: Cell cycle, cell division controls and transformation, growth factors, genes for social control of cell division, mechanism of cell division, cell adhesion, cell junctions and the extra cellular matrix, growth, development and differentiation.
	UNIT III
25 – 26	History of molecular biology
27 – 30	Nucleic acid as hereditary material, structure of DNA, chromatin, rRNA, tRNA and mRNA, proteins.
31 – 36	DNA replication, transcription, translation, genetic code, operon, positive and negative control of gene expression, important enzymes such as RNA replicase, reverse transcriptase, ligase, polymerase, ribozyme, etc.
	UNIT IV
37 – 40	Molecular mechanism of mutation.
41 – 44	Molecular organization of cell, structure of genomes, synthetic chromosomes.
45 – 48	RNA processing and alternative splicing, molecular biology of photosynthesis, nitrogen fixation and stress tolerance, development and differentiation and molecular evolution, RNAi and application.

Suggested Readings

Lewin B. 2008. Gene IX. Jones & Bartlett.

Primrose SB. 2001. Molecular Biotechnology. Panima.

TwymanRM.2003. Advanced Molecular Biology. Bios Scientific.

Applied Molecular Biology

Course No.	ABT 603	Title	Applied Molecular Biology
Credits	2 + 1=3	Semester	II

THEORY

Lecture Nos.	Topic
	UNIT I
1 – 2	Enzymes used in molecular biology and recombinant DNA research,
3 – 4	Gene identification, construction of gene libraries
5 – 6	Cloning and expression vectors
7 – 8	Gene mapping and DNA structure analysis
	UNIT II
9 – 11	Methods of DNA sequencing
12 – 13	synthesis of double stranded DNA and complementary DNA
14 – 16	cDNA library identification and enrichment of recombinant clones
	UNIT III
17 – 18	Methods for transfer of cloned DNA
19 – 20	Analysis and expression of recombinant DNA
21 – 22	Site directed DNA alterations and gene manipulations
23 – 24	Cloning in bacteria, yeast, plant and animal cells
	UNIT IV
25 – 27	Genetics of tumourogenic region of agrobacteria and its applications in agriculture, veterinary and medical sciences
28 – 30	biotechnology applications for production of high value and industrial products
31 – 32	safety aspects of genetic manipulations

PRACTICAL

Practical Nos.	Name of the experiment
1	Extraction of DNA
2	Extraction of RNA
3	Polyacrylamide gel electrophoresis (PAGE)
4	Agarose gel electrophoresis
5	Restriction endonuclease analysis of DNA.
6	Isolation and purification of plasmids
7	Polymerase chain reaction
8	Polymerase chain reaction contd.
9	Cloning of gene
10	Cloning of gene contd.
11	Expression of cloned gene
12	Expression of cloned gene contd.
13	Purification of recombinant protein
14	Blotting
15	RFLP
16	RAPD

Suggested Readings:

Kun LY. 2006. Microbial Biotechnology. World Scientific. Sambrook J & Russel DW. 2001. Molecular Cloning: a LaboratoryManual. ColdSpringHarbour Lab. Press.

TwymanRM.2003. Advanced Molecular Biology. Bios Scientific.

Animal Cell Cultures: Principle and Applications

Course No.	ABT- 604	Title	Animal cell cultures: Principle & applications
Credits	1+2 = 3	Semester	I

THEORY

Lecture No.	Name of Experiment
	UNIT I
1.	Introduction and Importance of cell culture, historical background of cell culture technology
2.	Tissue culture techniques – primary and secondary culture, continuous cell lines, suspension cultures, organ cultures etc
3.	Tissue culture techniques contd.
	UNIT II
4.	Different types of media for culturing cells and tissues
5.	Natural and defined media and growth supplements
6.	Culture of different tissues and its application
	UNIT III
7.	Behaviour of cells in terms of growth differentiation and metabolism
8.	Behaviour of cells contd.
9.	Cell division, growth patterns of cells
10	Estimation of cell number
	UNIT IV
11.	Development of cell lines
12.	Characterization and maintenance of cell lines
13	Characterization and maintenance of cell lines contd.
14	Stem cells and their applications
15	Cryopreservation
16	Common cell culture contaminants

PRACTICAL

Practical No.	Name of Experiment
1	Washing, preparation and sterilization of glassware for cell culture
2	Washing, preparation and sterilization of glassware for cell culture contd.
3	Preparation and sterilization of filter assembly
4	Filtration of media, trypsin, TVG solutions etc
5	Filtration of media, trypsin, TVG solutions etc contd.
6	Preparation of primary cell culture from chick embryo, kidney etc.
7	Preparation of primary cell culture contd.
8	Secondary culture of chick embryo fibroblast
9	Adaptation of cell lines to laboratory media
10	Adaptation of cell lines to laboratory media contd.
11	Subculturing and maintenance of cell lines
12	Subculturing and maintenance of cell lines contd.
13	Quantification of viable cells by trypan blue dye exclusion technique
14	Study the cytopathic effects of viruses on Primary cell cultures
15	Study the cytopathic effects of viruses on Primary cell cultures contd.
16	Study the cytopathic effects of viruses on cell lines
17	Study the cytopathic effects of viruses on cell lines contd.
18	Harvesting and preservation of viruses
19	Harvesting and preservation of viruses contd.
20	Study of effect of toxins/ toxic chemicals on cell lines
21	Study the effect of toxins/ toxic chemicals on cell lines contd.
22	Study the effect of drugs on cell lines
23	Study the effect of drugs on cell lines contd.
24	Isolation and cultivation of lymphocytes
25	Isolation and cultivation of lymphocytes contd.
26	Study of viral/bacterial internalization in lymphocytes
27	Study of viral/bacterial internalization in lymphocytes contd.
28	Suspension culture technique
29	Study of characteristics of suspension cell cultures
30	Preparation of cryoprotectant media
31	Cryopreservation of primary cell cultures
32	Cryopreservation of cell lines

Textbooks:

1. Freshney RI. 2005. Culture of Animal Cells. Wiley Liss.

2. Portner R. 2007. Animal Cell Biotechnology. Humana Press.

Molecular Diagnostics

Course No.	ABT-605	Title	Molecular Diagnostics
Credits	1 + 2 = 3	Semester	II

THEORY

Lecture No.	Topic
	UNIT I
1.	Introduction, importance and historical perspective of development of molecular diagnostic technology
2.	Polyacrylamide gel electrophoresis: Principle and applications
3.	Western Blotting: Principle and applications
4.	Concept of development of group specific and strain specific nucleic acid based diagnostics
5.	Basis for selection of gene/nucleotide sequence of pathogenic organism to target for detection
	UNIT II
6.	Application of restriction endonuclease analysis for identification of pathogens,
7	Principle of development of pathogen specific DNA probes
8.	Southern and Northern hybridization
	UNIT III
9.	Theoretical back ground of development of PCR
10	Different types of PCR assays
11.	Application of PCR for diagnosis of infectious diseases of animals and poultry
12.	Nucleic acid sequence based diagnostics
	UNIT IV
13	Advancements in diagnostic technology including DNA array technology
14	Advancements in diagnostic technology including DNA array technology contd.
15	Biosensors and Nanotechnology
16	OIE guidelines in development of diagnostics

PRACTICAL

Practical Nos.	Experiment
1.	Preparation of buffers and reagents
2.	Collection, preservation and transport of clinical and environmental samples from animal and poultry farms for molecular detection of pathogens
3 – 4	Isolation of bacterial pathogens from clinical specimens
	Isolation of bacterial pathogens contd.
4 – 5	Extraction of nucleic acids from bacteria and clinical specimens
6 – 7	Restriction endonuclease digestion and analysis by agarose gel electrophoresis
8 – 9	Development of animal pathogen specific nucleic acid probes
10 – 11	Southern blotting for animal pathogens
12 – 14	PCR for identification of bacterial pathogens from cultures
	PCR for detection of bacterial pathogens from blood
	PCR for detection of bacterial pathogens from animal tissues, clinical specimens
15 – 16	RT-PCR for detection of RNA viruses
17 – 19	Real time PCR for detection of pathogens in semen and other animal tissues
20 – 23	DNA fingerprinting for identification of animal species: Extraction of DNA from meat/tissue/ blood samples
	PCR amplification of mitochondrial 12 S rRNA gene
	Restriction enzyme digestion of the PCR product
	Analysis of RE digests by agarose gel electrophoresis and interpretation of results
24 – 26	PCR based detection of adulteration in processed and unprocessed meats: Extraction of DNA from meat samples
	PCR amplification
	Restriction fragment length polymorphism analysis
27 – 28	Detection of food borne pathogenic organisms in vegetables and fruits using PCR technology
29 – 30	PCR based detection of potential pathogens in milk, eggs and meat
31 – 32	Polyacrylamide gel electrophoresis and Western blotting

Suggested readings:

Elles R. and Mountford, R. 2004. Molecular diagnosis of genetic disease. Humana Press

Rao, R. R., fleming C. C. and Moore, J. E. 2006. Molecular Diagnostics. Horizon Bioscience.

Vaccine Biotechnology

Course No.	ABT-606	Title	Vaccine Biotechnology
Credits	2+1=3	Semester	III

THEORY

Lecture Nos.	Topic
	UNIT I
1	History of vaccinology
2 – 3	Conventional approaches to vaccine development
4 – 5	Live attenuated and killed vaccines
6 – 7	Preservation and monitoring of microorganisms in seed lot systems
8 – 9	Adjuvants and immunomodulators
	UNIT II
10 – 11	Instruments related to monitoring of temperature, sterilization, environment, quality assurance and related areas
12 – 13	Production techniques
14 – 15	Growing the microorganisms in maximum titre
16 – 17	Preservation techniques to maintain good antigen quality- Freeze drying, ultra freezing, etc.
18 – 19	Large scale production of vaccines and automation
	UNIT III
20 – 24	Recent developments in vaccine technology – subunit vaccines, synthetic vaccines, rDNA vaccines, idiotypic, DNA, glycoconjugate, deletion and edible vaccines, nanoparticles in vaccine delivery systems etc.
25 – 26	GMPs, quality control of conventional and recombinant vaccines
	UNIT IV
27 – 28	Introduction to pharmacopeal requirement
29 – 30	Disease security and biosecurity principles
31 – 32	OIE guidelines such as seed management, method of manufacture, in-process control, batch control, tests on final product

PRACTICAL

Practical No.	Experiment
1	Cultivation of virus in embryonated chicken eggs or bacteria on culture media
2	Harvesting of virus from inoculated eggs
3	Cultivation of bacteria on culture media
4	Harvesting of bacterial from inoculated culture media
5	Inactivation of harvested virus or bacterial cultures
6	Safety and sterility testing of inactivated vaccine
7	Safety and sterility testing contd.
8	Inoculation of tissue culture for propagation of virus
9	Harvesting and production of inactivated virus vaccine
10	Isolation and cloning of genes encoding immunogenic proteins
11	Isolation and cloning of genes encoding immunogenic proteins contd.
12	Expression of cloned gene
13	Expression of cloned gene contd.
14	Purification of recombinant immunogenic protein
15	Immunogenecity studies of recombinant protein: Immunization of laboratory animals or poultry
16	Immunogenecity studies contd.: Titration of antibodies against recombinant protein

Suggested Readings:

1. Barry R Bloom, Paul-Henri Lambert 2002. The Vaccine Book. Academic

Press.

2. Levine MM, Kaper JB, Rappuoli R, Liu MA, Good MF. 2004. New

Generation Vaccines. 3rd Ed. Informa Healthcare.

3. Lowrie DB & Whalen R. 2000. DNA Vaccines. Humana Press.

4. Robinson A & Cranage MP. 2003. Vaccine Protocols. 2nd Ed. Humana

Press.

Immunology Applied to Biotechnology

Course No.	ABT-607	Title	Immunology applied to Biotechnology
Credits	1+1=2	Semester	III

Lecture No.	Topic
	UNIT I
1.	Introduction to Immunology
2.	Principles of Immunology
3.	Immune system – organs and cells of the immune system
4.	Immune response – specific, nonspecific, passive, active responses
5.	Major Histocompatibility complex – different classes, comparative study of different species
6.	Various techniques used in biotechnology
	UNIT II
7.	Application of antibodies in purification of recombinant proteins
8.	Immunoblotting of recombinant proteins
9.	Expression of recombinant proteins
10.	Detection of recombinant proteins in different expression systems
11.	Industrial production of cytokines and interferons and purification
12.	Expression of immunoglobulin genes in plants and production of antibodies
	UNIT III
13.	Application of antibodies in chemiluminescence studies and fluorescence assays for recombinant genes
14.	Antibody based nucleic acid probes and their applications
15	Immunoinformatics
	UNIT IV
16.	Somatic cell hybridization, hybridoma technology, commercial production of monoclonal antibodies

PRACTICAL

Practical No.	Experiment
1.	Immunodiffusion
2.	Immunoelectrophoresis
3.	Fluorescent antibody test
4.	Immunoperoxidase test
5.	Enzyme linked immunosorbent assays (ELISA)
6.	PAGE
7.	Immunoblotting
8.	Separation of immunoglobulins from serum by ammonium sulphate pptation and dialysis
9.	Affinity chromatography
10.	Bioinformatics tools for immunological research
11.	Cultivation of normal lymphocyte culture
12.	Maintenance of myeloma cell line
13	Cell fusion and observation of cell clones
14	Identification of hybrid clones
15	Screening of hybrids for production of monoclonal antibodies and titration of antibodies from the antibody producing clones
16	Subculturing and maintenance of antibody producing clones.

Suggested Readings:

1. Barry R Bloom, Paul-Henri Lambert 2002. The Vaccine Book. Academic

Press.

2. Levine MM, Kaper JB, Rappuoli R, Liu MA, Good MF. 2004. New

Generation Vaccines. 3rd Ed. Informa Healthcare.

3. Lowrie DB & Whalen R. 2000. DNA Vaccines. Humana Press.

4. Robinson A & Cranage MP. 2003. Vaccine Protocols. 2nd Ed. Humana

Press.

Introduction to Bioinformatics

Course No.	ABT 608	Title	Introduction to Bioinformatics
Credits	1 + 1 = 3	Semester	III

THEORY

Lecture Nos.	Topic
	UNIT I
1	Introduction, biological databases – primary, secondary and structural
2	Protein and Gene Information Resources – PIR, SWISSPROT, PDB, genebank, DDBJ
3	Specialized genomic resources
	UNIT II
4 – 5	DNA sequence analysis,
6 – 7	cDNA libraries and EST, EST analysis
8	Pairwise alignment techniques
9	Database searching
10	Multiple sequence alignment
	UNIT III
11	Secondary database searching,
12	Building search protocol
13	Computer aided drug design – basic principles, docking, QSAR
	UNIT IV
14	Analysis packages – commercial databases and packages
15	GPL software for bioinformatics
16	Web-based analysis tools.

PRACTICAL

Practical No.	Experiment
1 – 3	The knowledge of various databases and bioinformatics tools available at NCBI the major content of the databases, purpose and utility
4 – 5	Open access bibliographic resources and literature databases
6 – 10	Primary and secondary databases, Retrieval of sequence / structure from databases: Nucleic acid sequence databases, Protein sequence databases Genome Databases, Visualization of structures
11 – 13	Docking of ligand receptors
14 – 16	Sequence-based Database Searches – BLAST exercises, sequence analysis using on-line tools, interpretation of results

Suggested Readings:

Attwood TK & Parry-Smith DJ. 2003. Introduction to Bioinformatics. Pearson Education.

Rastogi SC, Mendiratta N & Rastogi P. 2004. Bioinformatics: Concepts, Skills and Applications. CBS.

Animal Genomics

Course No.	ABT-609	Title	Animal Genomics
Credits	2+1=3	Semester	II

THEORY

Lecture Nos.	Topic
	UNIT I
1	Historical perspective
2 – 3	Genome organization in eukaryotes
4 – 5	Satellite DNA including mini and macro satellites and their various families, Long and short interspersed nucleotide elements
6 – 8	DNA markers – RAPD, STR, SSCP,RFLP, DNA fingerprinting, SNP, EST etc..
	UNIT II
9 – 10	Importance of gene mapping in livestock, methods and techniques used for gene mapping
11 – 12	Physical mapping, linkage analysis, cytogenetic techniques
13 – 15	FISH technique in gene mapping, somatic cell hybridization, radiation hybrid maps, in-situ hybridization, comparative gene mapping
	UNIT III
16	Genetic distance analysis
17 – 18	Breed characterization on the basis of DNA markers
19 – 20	Genetic markers for quantitative traits loci
21 – 22	Marker assisted selection for incorporation of desirable traits DNA markers with economic traits
23 – 24	Restriction fragment length polymorphism (RFLP) of different structural genes
	UNIT IV
26 – 27	Current status of gene maps of livestock
27 – 28	MHC and its relevance to disease resistance and immune response
29 – 30	Genes influencing production traits
31 – 32	Mitrochondrial DNA of farm animals, evolutionary significance, application of genome analysis of animals in breeding

PRACTICAL

Practical No.	Experiment
1.	Preparation of lymphocyte culture from cattle, buffalo etc.
2.	Fixation of cells and chromosomal preparation
3.	Karyotyping and observation of different banding patterns
4.	Isolation and purification of animal genomic DNA from blood lymphocytes from cattle, buffalo etc.
5.	Analysis of DNA by agarose gel or polyacrylamide gel electrophoresis
6.	Checking quality and quantity of genomic DNA by AGE
7.	Restriction digestion and analysis
8.	Southern hybridization
9.	DNA fingerprinting
10.	Techniques for detecting polymorphism – RAPD
11.	RFLP
12.	SSCP
13	AFLP
14	STRP
15	Cloning of genomic DNA or cDNA cloning
16	Differentiation of tissues of different species by mitochondrial genome analysis

Suggested Readings:

Gibson, G. and Muse S. V. 2004. A primer of Genome Science, Sinauer Associates

Primrose, S. B. and Twyman R. M. 2007. Principles of Genome analysis and genomics. Blackwell

Sensen C.W. 2005. Handbook of Genome Research. Vols. I, II. Wiley-CVH

Reproductive Biotechnology

Course No.	ABT 610	Title	Reproductive Biotechnology
Credits	2 + 1=3	Semester	III

THEORY

Lecture Nos.	Topic
	UNIT I
1 – 4	History, importance of assisted reproductive biotechnology in man animals, Introduction to embryo biotechnology, endocrine therapeutics
	UNIT II
5 – 10	Biotechnological approaches to reproduction: Applications, Advantages & disadvantages of ETT; Selection of donors and recipients for embryo transfer, Methods of synchronization of oestrus, Methodology of super ovulations, Methods of Embryo collection & embryo evaluation, Methods of embryo transfer
11 – 16	In vitro maturation (IVM), Preparation of semen, ,In vitro capacitation of sperm (IVC), In vitro fertilization(IVF), In vitro culture, Embryo culture and micromanipulation
	UNIT III
17 – 22	Different method of gene transfer and their limitations, embryo splitting, embryo sexing by different methods, production of transgenic livestock by nuclear transfer and its application, regulatory issues.
	UNIT IV
23 – 28	Cloning of domestic animals. Conservation of endangered species, Characterization of embryonic stem cells. Different applications of embryonic stem cells
29 – 32	Ultrasound techniques in pregnancy diagnosis, Laparoscopic techniques in embryo transfer

PRACTICAL

Practical Nos.	Name of the experiment
1	Preparation of glassware/plastic ware/reagents for ETT
2	Selection of donors and recipients
3 – 4	Synchronization and superovulation protocols
5	Collection of embryos using non-surgical procedures
6	Transferring embryos using non- surgical procedures
7	Oocyte collection and evaluation
8 – 9	In vitro maturation (IVM), Preparation of semen, In vitro capacitation of sperm (IVC)
10 – 11	In vitro fertilization (IVF)
12 – 13	Culture of embryos
14	Micromanipulation of early embryos
15	Embryo-spliting
16	Cryopreservation of embryos

Suggested Readings:

Ball P.J.H. and Peter A.R. 2004 Reproduction in cattle. Blackwell

Gordon,I.2003 Laboratory Production of Cattle Embryos. CABI

Gordon,I.2005 Reproductive Techniques in Farm Animals. CABI

Techniques in Molecular Biology & Genetic Engineering

Course No.	ABT 611	Title	Techniques in Molecular Biology & genetic Engineering
Credits	0 + 3 = 3	Semester	II

THEORY

Practical Nos	Experiment
	UNIT I
1 – 2	Preparation of solutions, reagents required for isolation of DNA and other molecular biology work
3 – 4	Isolation of bacterial plasmids
5 – 6	Isolation of chromosomal DNA from bacteria
7 – 8	Isolation of DNA from mammalian cells
9 – 10	Isolation of mRNA/RNA
11 – 12	Quantification of nucleic acids
	UNIT II
13 – 15	Plasmid minIprep; Isolation of plasmid and restriction endonuclease digestion
16 – 18	Restriction endonuclease digestion of chromosomal DNA
19 – 21	Agarose gel electrophoresis of RE digested DNA
22 – 24	cDNA synthesis
	UNIT III
25 – 26	Polymerase Chain Reaction using random primers
27 – 28	Polymerase Chain Reaction using specific primers
29 – 34	Different types of PCR, Multiplex PCR, Nested PCR, Real time polymerase chain reaction
	UNIT IV
35 – 37	Cloning of bacterial and viral genes in to plasmid vectors
38 – 40	DNA ligation and transformation
41 – 44	Confirmation of insert by RE digestion and touch PCR
45 – 48	Synthesis of nucleic acid probes, Nucleic acid hybridization

Suggested Readings:

Kun LY. 2006. Microbial Biotechnology. World Scientific.

Sambrook J & Russel DW. 2001. Molecular Cloning: a LaboratoryManual.ColdSpringHarbour Lab. Press.

TwymanRM.2003. Advanced Molecular Biology. Bios Scientific.

Molecular Forensics

Course No.	ABT 613	Title	Molecular Forensics
Credits	2 + 1 = 3	Semester	III

THEORY

Lecture Nos.	Topic
	UNIT I
1	General history of forensic science
2	introduction to DNA forensics
3 – 4	Scope and application of DNA forensics in animal and human criminal investigations in variety of situations
	UNIT II
5 – 6	Isolation of nucleic acids and quantification
7 – 8	DNA finger-printing: Different methods
9 – 10	PCR and its applications
11 – 12	Nucleic acid hybridization
13 – 14	Restriction endo-nuclease analysis and sequencing
15 – 16	Mitochondrial DNA analysis and its applications
17 – 18	Individual Animal Identification using DNA fingerprinting
	UNIT III
19 – 21	Animal species identification in religious disputes
22 – 24	Identification of adulteration of meat by DNA based techniques
25 – 27	Molecular techniques for investigations into theft of farm animals, pets etc.
28	Advantages, disadvantages and limitations of DNA forensics
	UNIT IV
29 – 30	Mass spectroscopy for characterization of proteins
31 – 32	Immunological techniques viz. ELISA, immunoelectrophoresis and immunofluorence.

PRACTICAL

Practical Nos.	Name of the experiment
1	Collection, preservation, dispatch and storage of material for forensic analysis
2	Preparation of solutions, reagents required for forensic analysis employing molecular techniques
3	Isolation of nucleic acid from blood, milk and other body fluids
4	Isolation of nucleic acid from wild animal scat
5	Isolation of nucleic acid from skin, meat, hair, cooked and putrefied meat
6	Designing of primers
7	Polymerase chain reaction (PCR) for identification of species
8	Multiplex PCR for species identification
9	Random amplified polymorphic DNA (RAPD)
10	Restriction fragment length polymorphism (RFLP) analysis
11	Identification of meat animal species by PCR RFLP analysis of mitochondrial 12S rRNA gene
12	Identification of meat animal species by PCR RFLP analysis contd.
13	Detection of adulteration in meat by PCR
14	Detection of adulteration in meat by nucleic acid hybridization assay
15	Detection of adulteration in meat contd.
16	Immunological techniques for identification of meat animal species

Suggested Readings:

Lincoln PJ & Thomson J. 1998. Forensic DNA Profiling Protocols. Humana Press.

Rudin N & Inman K. 2002. An Introduction to Forensic DNA Analysis. 2^nd Ed. CRC Press.

Animal Biotechnology

Course No.	ABT 616	Title	Animal Biotechnology
Credits	3 + 0 = 3	Semester	II

THEORY

Lecture Nos.	Topic
	UNIT I
1	Structure of animal cell , History of animal cell culture
2 – 3	Cell culture media and reagents, culture of mammalian cells, tissues and organs, primary culture, secondary culture, continuous cell lines, suspension cultures
4	Somatic cell cloning and hybridization, transfection and transformation of cells
5	Commercial scale production of animal cells
6	Application of animal cell culture for in vitro testing of drugs
7	Testing of toxicity of environmental pollutants in cell culture
8	Application of cell culture technology in production of human and animal viral vaccines and pharmaceutical proteins
	UNIT II
9	Introduction to immune system
10	Cellular and humoral immune response
11	History of development of vaccines, introduction to the concept of vaccines
12	Conventional methods of animal vaccine production
13	Recombinant approaches to vaccine production,
14	Hybridoma technology
15 – 16	Phage display technology for production of antibodies
17 – 18	Antigen-antibody based diagnostic assays including radioimmunoassays and enzyme immunoassays, immunoblotting,
19 – 20	Nucleic acid based diagnostic methods
21	Commercial scale production of diagnostic antigens and antisera, animal disease diagnostic, kits, probiotics.
	UNIT III
22	Structure of sperms and ovum
23	Cryopreservation of sperms and ova of livestock, artificial insemination
24 – 25	Super ovulation, in vitro fertilization, culture of embryos, cryopreservation of embryos, embryo transfer, embryo-spliting, embryo sexing
26 – 27	Transgenic manipulation of animal embryos, different applications of transgenic animal technology,
28	Animal cloning basic concept, cloning from- embryonic cells and adult cells,
29	Cloning of different animals, cloning for conservation for conservation endangered species
30	Ethical, social and moral issues related to cloning
31	In situ and ex situ preservation of germplasm
32	In utero testing of foetus for genetic defects, pregnancy diagnostic kits, anti-fertility animal vaccines,
33	Gene knock out technology and animal models for human genetic disorders
	UNIT IV
34 – 35	Introduction to different breeds of cattle, buffalo, sheep, goats, pigs, camels, horses, canines and poultry
36 – 37	Genetic characterization of livestock breeds,
38	Marker assisted breeding of livestock,
39	Introduction to animal genomics,
40 – 41	Different methods for characterization of animal genomes, SNP, STR, QTL, RFLP, RAPD
42	Genetic basis for disease resistance,
43	Transgenic animal production and application in expression of therapeutic proteins
44	Immunological and nucleic acid based methods for identification of animal species
45	Detection of meat adulteration using DNA based methods
46	Detection of food/feed adulteration with animal protein
47 – 48	Identification of wild animal species using DNA based methods using different parts including bones, hair, blood, skin and other parts confiscated by anti-poaching agencies

Suggested Readings:

GordonI.2005. Reproductive Techniques in Farm Animals. CABI

Kindt TJ, Goldsby RA & Osbrne BA. 2007. Kuby Immunology. WH Freeman

Kun LY. 2006. Microbial Biotechnology. World Scientific

Levine MM, Kaper JB, Rappuoli R, Liu MA, Good MF. 2004. New Generation Vaccines. 3rd Ed. Informa Healthcare

Lincoln PJ & Thomson J. 1998. Forensic DNA Profiling Protocols. Humana Press.

Portner R. 2007. Animal Cell Biotechnology. Humana Press.

Spinger TA. 1985. Hybridoma Technology in Biosciences and Medicine. Plenum Press.

TwymanRM.2003. Advanced Molecular Biology. Bios Scientific.