BIOTECHNOLOGY (878)

Aims:
1. To enable candidates to acquire the knowledge 4. To create awareness about the appreciation of
and develop an understanding of how materials biological processes to industries.
are provided by biological agents to provide 5. To develop the ability to appreciate biological
goods and services. phenomenon in nature and the contribution of
2. To appreciate the role played by biotechnology biotechnology to human welfare.
in improving health care for human beings. 6. To develop scientific attitude towards biological
3. To understand the interdisciplinary nature of this phenomenon.
subject.
CLASS XI
There will be two papers in the subject: of microorganisms involved in their
production.
Paper I: Theory…………... 3 hours ... 70 marks
Application of these technologies for large-
Paper II: Practical………. 3 hours ... 15 marks
scale production, with special reference to
Project Work………. … 10 marks fermentation (Beer production only). Quality
Practical File…………. … 5 marks control management of the products, good
laboratory practices.
PAPER I –THEORY- 70 Marks (ii) Scope and importance of biotechnology:
There will be one paper of three hours duration different branches of biotechnology and
divided into two parts. different regulatory guidelines; ethical, legal
and social issues (ELSI) that a biotechnologist
Part 1 (20 marks) will consist of compulsory short comes across while doing the work. Various
answer questions, testing knowledge, application and organisations in the field of biotechnology.
skills relating to elementary/fundamental aspects of
the entire syllabus. Names, definitions and importance of various
fields that can be covered under
Part 2 (50 marks) will consist of eight questions out biotechnology such as - agricultural/ plant
of which the candidates will be required to answer biotechnology, animal biotechnology/medical
five questions. Each question in this part shall carry biotechnology, nanobiotechnology, industrial
10 marks. biotechnology, immunology and health care,
1. Introduction to Biotechnology energy and environment.

(i) Historical background; definition; a brief Intellectual Property Rights (IPRs) in

introduction of the traditional and modern biotechnology- concept of intellectual
techniques of Biotechnology and their property, types of IPR and its need;
applications. intellectual property rights and the choice of
intellectual property rights protection.
Definition of biotechnology by OECD and Discovery and invention; Concept of
EFB; contributions of Karl Ereky and Louis patenting, trademark, trade secrets,
Pasteur; use of various fermented products in copyright, geographical indications and PBRs
ancient civilisations; and their need.
Kitchen (traditional), the first Concept of ethical, legal and social issues
biotechnological laboratory -reasoning with one common example IVF.
behind the technology involved in simple
biological products like curd and beer; names

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 Biosafety issues: release of genetically chromatin and nucleosome. An idea about
modified organisms into the environment and banding patterns (Q, R, C and G) and their
their impact; GEAC and its objectives. application.
Biotechnology - global and Indian scenario. Concept of chromosomal number in different
Various institutes, centres and funding species, e.g. man, mouse, Drosophila and pea.
agencies - NBTB, CCMB, ICGEB, ICMR,
Techniques in cytology – microscopy (light
ICAR, DBT, DST which deal with
and electron microscope), karyotyping and
biotechnology and bioinformatics in India:
centrifugation (principle and applications
names only.
only).
(iii) Basic concepts of Biochemical technology
(ii) Cell Division and cell cycle: types of cell
and biostatistics: What does the biochemical
divisions and various other activities of cell
technology mean? An understanding of
such as biochemical transformations.
various statistical methods involved in
biotechnology. Types and significance of cell division and a
brief note about the different stages of cell
Concept of buffer, type and preparation of
division – mitosis and meiosis.
buffers, pH, physical variables; fermentation;
An understanding of bio-reactors, idea of Basic concept of cell cycle and cell cycle
sampling – quadrat and transect; measures of regulation – CdK method only, definition of
central tendency – mean, median, mode; Mitotic Index.
standard deviation and standard error; Biochemical Transformations:
concept of probability – theoretical and
experimental. An understanding of biochemical
transformations, different biochemical
2. Cell Biology pathways involved in respiration - aerobic
(i) Cell: Justification of cell as a basic unit of and anaerobic.
life. Prokaryotic cell and eukaryotic cell; A Aerobic respiration - Glycolysis, Krebs’ cycle,
brief note on the cell components with special electron transport chain and oxidative
reference to nucleus. Various cytological phosphorylation.
techniques used in identifying the cell and
chromosomes. Anaerobic respiration - lactic acid,
fermentation and alcohol fermentation –
Differentiation prokaryotic and eukaryotic definition only.
cellular systems.
(iii) Errors in cell division: what happens if the
Structure of bacteria (in brief, with reference cell does not divide normally? An
to plasmid). Gram+ and Gram- bacteria. understanding of different numerical and
An understanding of cell components, their structural abnormalities.
basic structure and functions - cell wall, cell Concept of mutation: causes; types –somatic,
membrane, cytoplasmic reticulum, Golgi germinal, spontaneous, induced, gene,
apparatus, mitochondria, ribosomes, chromosomal and genomatic mutations,
vacuoles, plastids, lysosomes, nucleus and euploidy, aneuploidy, monosomy, nullisomy,
other important inclusions of the cell. trisomy and tetrasomy; various factors
Chromosomal structure and composition – causing mutations.
organisation of chromatids, concept of Concept of non-disjunction: meiotic
homologous and non-homologous non-disjunction and mitotic non-disjunction.
chromosomes, sister and non-sister Non-disjunction in sex chromosomes –
chromatids, classification of chromosomes on Turner’s syndrome and Klinefelter’s
the basis of position of the centromere on the syndrome - chromosomal composition and
chromosome, basic idea about telomere, symptoms only.
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 Numerical chromosomal aberrations with energy; mechanism of enzyme action - lock and
respect to autosomes, i.e. Down’s syndrome – key model; induced fit hypothesis; factors
chromosomal composition and symptoms affecting enzyme activity (temperature, pH,
only. substrate concentration, enzyme concentration,
inhibitors (competitive, non-competitive).
Structural chromosomal abnormalities –
deletions, duplications, translocations, Optical activity of biomolecules
inversions. (dextrorotatory and laevorotatory).
Polyploidy and its significance in plants. Concept of supramolecular assembly.
Inborn errors of metabolism - basic concept (ii) Techniques used for separation of
and examples like albinism, sickle cell biomolecules
anaemia, phenylketonuria and alkaptonuria.
Ion exchange chromatography and paper
chromatography.
3. Biomolecules and related techniques
(i) Introduction to biomolecules- definition and 4. Developmental Biology and Immunology
types. Carbohydrates, proteins, lipids, vitamins
(i) Animal and plant development: development
and enzymes – their structure and properties.
of an organism from zygotic cell in both plants
Biomolecules – definition and types and animals.
Structure and functions of carbohydrates. Animal development – fertilisation, zygote to
blastocyst formation.
Sugars and derivatives; classification of some
important mono, di and polysaccharides - Plant development. Double fertilisation
glucose, fructose, glycogen, cellulose, chitin including formation of primary endosperm
and peptidoglycan. Physical and chemical nucleus.
properties of sugars.
(ii) An understanding of defence strategies in
Structure, functions and classification of living organisms.
proteins i.e. simple, complex and derived;
Immune system in higher animals, concept of
building blocks of proteins - the amino acids:
immunity, immunisation, antigen and antibody.
chemical structure, types (acidic, basic and
Various cells involved in immune response in
neutral); physical and chemical properties of
humans. An introduction to human leukocyte
amino acids. 3D - structure of proteins.
antigens with reference to organ
Different types of protein structures - primary,
transplantation; Types of immunity - innate
secondary (alpha helix, beta pleated sheet and
and acquired. ELISA Technique (Enzyme
random structures), tertiary, quaternary;
Linked Immuno Sorbent Assay).
protein sequencing by MALDI-MS.
Secondary metabolites in plants and their
Structure and functions of lipids – fatty acids
significance
and alcohol; types (simple, conjugated and
derived lipids with one example of each); Defence strategies in bacteria – endospores
chemical and physical properties of lipids. and R plasmids.
Vitamins: Definition, types (fat soluble and 5. Genetics
water soluble vitamins); co-enzyme forms of
water soluble vitamins; deficiency diseases of (i) Laws of Inheritance: An account of Mendel’s
vitamins. experiments. Different types of genetic
inheritance.
Enzymes: Structure and functions of enzymes:
chemical nature of enzymes; characteristics Mendel’s experiment on pea plant and his
and properties of enzymes. An understanding laws of inheritance.
of enzyme activity on the basis of activation

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 Concept of trait, gene, allele, phenotype, once and destroy it. (d) Do not prick or use blood
genotype, homozygosity, heterozygosity and drop in an indiscriminatory way.
hemizygosity. Types of inheritance:
2. Identification of different types of blood cells by
autosomal inheritance - dominant, co-
preparing blood smear using Leishmann’s stain.
dominant, recessive, polygenic, pleiotropic
and cytoplasmic inheritance (plastidial Requirements: Blood sample, disposable needles,
inheritance). slides, Leishmann’s stain. Make a blood smear on
a slide, use the stain to colour the smear, wash
Pedigree construction using different
and observe under microscope.
standard symbols.
Sex chromosome inheritance - with special 3. Instruments – their names, use and principles (if
reference to X chromosomal inheritance with applicable).
suitable examples (colour blindness and Water bath, pH meter, weighing balance,
haemophilia). desiccators, microfiltration unit, magnetic stirrer,
(ii) Gene Mapping: mapping of genes on LAF, haemocytometer, micropipette, vortex mixer,
chromosomes using linkage analysis. Cancer colorimeter/spectrophotometer, hot air oven,
and its genetics. autoclave, incubator, electrophoresis chamber,
colony counter, autoclave, hot plate.
Mapping of genes on chromosomes with
respect to COV (Crossing Over Value). 4. Finding out the pH of water by using pH meter or
Basic concept of linkage (types not required) pH paper on tap water and water containing acid,
and crossing over. Genetic recombination. base.
Cancer: Causes (physical, chemical, Take tap water in three test tubes, add two drops
biological – TSG and oncogenes); diagnosis of dil. HCl in one, two drops of NaOH in the
and treatment. second while leaving the third test tube with tap
water. Use pH meter or pH paper to find their
(iii) Genes in populations: how do genes behave
specific pH.
in populations from generation to
generation? Various ways of studying 5. Observation of steps of mitosis by using the root
tip of onion.
population genetics.
The students should be given practice in
Concept of gene pool and allele frequency, preparing slides for study of mitosis by crush
definition of Hardy Weinberg law, its smear method. They should be able to identify
applications. different stages (at least four stages). The
Possibility of disease resistant and requirement for this set of experiments is
susceptible genes in population. Definition Acetocarmine stain slides, coverslips,
and application of pharmacogenetics and microscopes and spirit-lamp.
pharmacogenomics.
6. Measurement of mitotic index.
PAPER II Mitotic index is the ratio of number of cells
undergoing mitosis to the number of cells in the
PRACTICAL WORK – 15 Marks field.
Candidates are required to complete the following
experiments. No. of cells showing mitosis
MI =
1. Determination of blood group by using antisera. Total no. of cells in the field
The students can perform this experiment on their 7. Observation of various stages of meiosis under
own blood groups. Proper instructions however microscope.
are to be given for ‘prick’ – e.g. (a) Sterilize For the study of meiosis, the students should be
finger with alcohol/disinfectant. (b) Use only shown permanent slides of meiosis and they
disposable sterile needle. (c) Use the needle only should be able to identify at least six stages of
meiosis from the slides.

304
8. Effect of temperature on curdling of milk by using milk is mixed with water, it would sink higher
Lactobacillus bacteria at 37oC, 60oC and 10oC. than mark ‘M’. If it stands at the mark 3 it
Optimum temperature for curdling of milk is 37oC means that the milk is 75% pure and
due to active form of bacteria at this temperature; respectively 2 for 50% purity and 1 for 25%
it is inactive at low temperature and dies at high purity.
temperature. 11. Construction of pedigree showing different
9. Food tests: types of inheritance.
(i) Carbohydrates – starch by iodine solution
The students are to observe the traits like,
turning blue - black in colour.
rolling of the tongue/ attached earlobe/
Reducing and non-reducing sugars by using widow’s peak.
Fehling’s solution / Benedict’s solution –
reducing the cupric ion (blue) to cuprous ion 12. Preparation of karyotypes.
(red). Demonstration of any metaphasic plate of
(ii) Protein test – Biuret test, Xanthoproteic mitosis.
and Millon’s test 13. Sampling methods – quadrat and transect by
(a) For Biuret test –The protein produces using different techniques.
deep blue – violet colour due to the
involvement of cupric ion in the product To be done in groups. Use yellow and green
formed. pea seeds. Make a quadrat (30 cm X 30 cm)
with blocks of 6 cm X 6 cm. Spread the seeds
(b) For Millon’s Reagent – A pinkish red
randomly on the table top. Put the quadrat
colour is observed with mercuric and count the number of yellow and green
chloride.
peas per block; find the frequency of each
(c) For Xanthoproteic Test: When type of pea seed.
concentrated nitric acid is boiled with
protein a yellow colour is observed. On 14. Data collection – primary and secondary.
addition of ammonium hydroxide or Collect any type of primary data and
liquor ammonia orange yellow precipitate secondary data, tabulate the data and draw
is obtained. conclusion.
(iii)Lipids – Sudan III, Acrolein test, paper test
PROJECT WORK AND PRACTICAL FILE
(a) Sudan III is a red fat-soluble dye used for
identification of the presence of lipids, – 15 marks
triglycerides and lipoproteins. It reacts Project Work – 10 Marks
with the lipids or triglycerides and gives Candidates are to creatively execute one
red colour. project/assignment on any aspect of Biotechnology.
(b) Acrolein test is used to detect fat. When Teachers may assign or students may choose any one
fat is heated strongly in the presence of project of their choice. The report should be kept
potassium bisulphate/ sodium bisulphate simple, but neat and elegant. No extra credit shall be
(KHSO 4 /NaHSO 4 ) that acts as a given for type-written material/decorative cover, etc.
dehydrating agent, the glycerol is
Practical File – 5 Marks
dehydrated to form an unsaturated
aldehyde called acrolein that gives a Teachers are required to assess students on the basis
pungent and irritating odour. of the practical file maintained by them during the
academic year.
10. Finding out the purity of milk by using
lactometer.
Put the instrument in milk. If it sinks down
and reaches the mark ‘M’ mentioned on
lactometer, it means that the milk is pure or if
not, it means that the milk is impure. If the
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 LIST OF ABBREVIATIONS
1. CCMB:Centre for Cellular and Molecular Biology 15. ICGEB: International Centre for Genetic
Engineering and Biotechnology
2. CdK: Cyclin dependent Kinase
16. ICMR: Indian Council for Medical Research
3. COV: Cross Over Value
17. IEF : Iso Electro Focussing
4. CSIR: Council of Scientific and Industrial
Research 18. IPP: Intellectual Property Right Protection Act

5. DBT: Department of Biotechnology 19. IPR: Intellectual Property Right

20. IVF: In–Vitro Fertilization
6. DST: Department of Science and Technology
21. MALDI-MS: Matrix Assisted Laser Desorption
7. EFB: European Federation of Biotechnology Ionization – Mass Spectrometry
8. ELISA: Enzyme Linked Immuno Sorbent Assay 22. MI: Mitotic Index
9. ELSI: Ethical, Legal and Social Issues 23. NADPH/NADP: Nicotinamide Adenine
10. ETS/ETC: Electron Transport System / Electron Dinucleotide Phosphate (reduced) / Nicotinamide
Transport Cycle Adenine Dinucleotide Phosphate

11. FMN/FAD: Flavin Mono Nucleotide / Flavin 24. NBTB: National Biotechnology Board
Adenine Dinucleotide 25. OECD: Organization for Economic Cooperation
and Development
12. GEAC: Genetic Engineering Approval Committee
26. PBR: Plant Breeder’s Right
13. HLA: Human Leucocyte – associated Antigen
27. TPP: Thiamine Pyrophosphate
14. ICAR: Indian Council for Agricultural Research
28. TSG: Tumour Suppressor Gene

306
 CLASS XII
There will be two papers in the subject: Techniques of nucleic acid estimation –
colorimetry and UV-visible
Paper I: Theory……………. 3 hours ... 70 marks
spectrophotometry.
Paper II: Practical…………. 3 hours ... 15 marks
(ii) Protein Synthesis: synthesis of different
Project Work………. … 10 marks RNAs, and the complete mechanism of
Practical File………… … 5 marks polypeptide chain formation.
Concept of central dogma.
PAPER I: THEORY- 70 Marks
From genes to proteins:
There will be one paper of three hours duration
divided into two parts. (a) Concept of transcriptional unit, promoter,
structural and terminator region; concept
Part 1 (20 marks) will consist of compulsory short of split gene - intron and exon;
answer questions, testing knowledge, application and monocistronic and polycistronic RNA,
skills relating to elementary/fundamental aspects of hnRNA;
the entire syllabus.
(b) Transcription – explanation of the
Part 2 (50 marks) will consist of eight questions out complete process including enzymes
of which the candidates will be required to answer involved in the process; Post-
five questions. Each question in this part shall carry transcriptional changes and their
10 marks. significance in eukaryotes –
1. Molecular Biology polyadenylation, capping and RNA
splicing;
(i) Nucleic acids and their estimation: an
understanding of nucleic acids, their (c) Concept of reverse transcription;
biochemical structure. (d) Genetic code – properties of genetic code,
DNA as the genetic material (Hershey and start and stop codons, anticodons.
Chase experiment). (e) The translation of RNA to protein –
DNA (B-DNA)– physical and chemical complete mechanism of chain initiation,
structure; definition, double helical model of elongation and termination, the role of
DNA, (Watson and Crick’s); Nucleotide and tRNA, mRNA and rRNA in protein
nucleoside; Chargaff’s Law, method of synthesis. (Post translational changes not
replication of DNA, various replicative included).
enzymes in both prokaryotic and eukaryotic (iii) Gene regulation in prokaryotes
organisms, example topoisomerases, helicase,
SSBs polymerases, primases, ligases. Concept Operon concept – lac operon and trp operon.
of semi conservative (with respect to 2. Genetic Engineering
Messelson and Stahl experiment and Taylor
(i) Introduction to gene cloning and genetic
et.al experiment on Vicia faba using
engineering: concept of cloning and vectors.
radiolabelled thymidine) and semi-
discontinuous replication, (leading and Tools of recombinant DNA technology, types of
lagging strands), okazaki fragments. restriction endonucleases and other enzymes
used in gene cloning; techniques involved in
RNA – definition, various types of RNAs such
extraction and purification of DNA from
as mRNA, tRNA (Clover leaf model with
bacterial, plant and animal cells.
diagram; brief introduction to L-shaped
model), rRNA their structure and functions. Selection of host cells: eukaryotic and
prokaryotic.

307
 Vectors: Characteristics and types such as (iii) Gene analysis techniques: various techniques
plasmids -pBR322, pUC (in pBR322- presence involved in recombinant DNA technology.
of two antibiotic resistant genes and in
pUCpresence of lac Z gene to be taught), DNA probes – definition and use.
cosmids, phages (M13 and λ), YACs, BACs (to Low resolution mapping techniques: gel
be taught with reference to stability and their electrophoresis, southern blotting (details of
carrying capacity), animal and plant viruses the technique to be taught), western and
(CaMV, retrovirus, SV40 – only names of northern blotting (a brief idea and their uses).
viruses, no details).
High resolution techniques: DNA sequencing-
Transfer of recombinants into host cells – sequencing by chain termination, automated
(a) Vectorless methods - basic concept of DNA sequencing. Site directed mutagenesis.
transformation, transfection, DNA amplification by Polymerase chain
electroporation, liposome mediated gene reaction (PCR)– applications of PCR, steps
transfer, microinjection, biolistic and application of DNA profiling or DNA
(b) Vector-mediated method - Agrobacterium finger printing.
tumefaciens induced gene transfer. 3. Cell culture technology
Methods of identification of recombinants- A brief idea of tools and techniques involved in
Direct selection (green fluorescent cell culture technology and their applications in
selection) and Insertional inactivation microbial, plant tissue and animal cell cultures
(Blue-white selection, antibiotic resistance). respectively.
A basic understanding of DNA libraries – (i) General tools and techniques used in cell
construction of genomic and cDNA culture technology
libraries.
(a) Instruments - centrifuge, LAF hood and
Construction of a recombinant DNA biosafety cabinets, pH meter, autoclave,
molecule. vortex mixer, hot air oven, magnetic
(ii) Innovations in Biotechnology: produced by stirrer, weighing balance, micro
using modern biotechnological tools, (select filtration unit, incubator, CO 2
incubator, inverted microscope,
examples of products already available)
bioreactor (diagram, its components and
(a) Plants: Production of Flavr Savr their function)-stirred tank and sparged
tomatoes, Bt-crops and Golden rice. type (brief idea only), use of T flasks to
propagate animal cells.
(b) Healthcare: Production of recombinant
hepatitis-B vaccine, Humulin, interferon Only uses of the above instruments to
and edible vaccines. be studied.
(c) Animal: Dolly the cloned sheep, Sources (b) Sterilization techniques for culture room,
and characteristics of stem cells and apparatus, transfer area, media,
their applications. vitamins, and living material;
(d) Environmental biotechnology: (c) Cryopreservation (need and steps).
bioremediation using oil-eating bacteria (d) Cell counting (direct counting by
as an example. haemocytometer), cell viability by
(e) Industrial biotechnology: applications of Evan’s blue stain and cell sorting (FACS
industrial enzymes – rennet, subtilisin, only)
amylase, papain. (e) Types of media (synthetic /defined, semi-
synthetic/differential, complex/natural)

308
 Preparation of media: microbial media- (iv) Animal cell culture and its application.
LB agar and LB broth; Plant media-MS Primary cell culture with mechanical and
and White’s media; Animal media- enzymatic disaggregation and its drawbacks;
RPMI, DMEM and FBS - brief idea only. Types of cell-lines: finite, continuous,
adherent and suspension; scale up-mono
(includes inorganic and organic layer by Roller bottle, application of animal
macronutrients and micronutrients, cell culture-tissue, hybridoma technology,
antibiotics, growth regulators for plants: tissue engineering (definition only).
auxins and cytokinins). 4. Bioinformatics
Importance of pH and solidifying agents. (i) Introduction to bioinformatics; global
bioinformatics databases and data retrieval
(ii) Microbial culture and its application. tools; genomics, different types of sequences,
Fermentation process and growth kinetics- types of sequence analysis.
batch culture, fed batch culture, continuous Introduction to bioinformatics: definition and
culture (with the help of graphs only): need.
Definition of turbidostat and chemostat: An introduction to global bioinformatics
Products and application-SCP (definition and databases (nucleotide and protein databases).
use), industrial enzyme-subtilisin (source and Information sources such as EMBL, NCBI,
its use). DDBJ, SWISSPROT, GenBank, GENSCAN.
Data retrieval tools- ENTREZ, Taxonomy
(iii) Plant tissue culture and its application.
Browser.
Isolation of single cell by mechanical and (ii) Genomics: Definition, introduction, tools used
enzymatic methods, synchronisation of cell in Genomics and its applications.
culture by chemical methods like starvation, Definition of genomics. Types of genomics-
inhibition and mitotic arrest. structural and functional. Basic criteria in
Cellular totipotency-definition of cellular selecting the organism for its genome
differentiation, de-differentiation, re- sequencing. Different types of sequences –
differentiation. Application of plant cell cDNA, genomic DNA, ESTs (Expressed
culture technology (methodology not Sequence Tags) and STSs (Sequence Tagged
required, only brief idea needed): Sites) and the different softwares (example
gene scan).
(a) Haploid production-androgenesis and
Types of sequence analysis by using BLAST
gynogenesis and their significance.
and FASTA –global, local, pair wise and
(b) Triploid production-understanding and multiple.
need for triploid production and its Human Genome Project - its objectives, the
application (seedless crops). countries involved, its achievements and
(c) In-vitro pollination- concept and its significance.
application. DNA microarray technology – definition and
(d) Zygotic embryo culture- concept and its application only.
application, Embryo rescue (brief idea Concept of Single Nucleotide Polymorphisms
only). (SNPs).
(e) Somatic hybridisation-protoplast fusion (iii) Proteomics: definition, introduction and
(Pomato). databases.
(f) Micropropagation and its significance. Types of Proteomics – structural, functional
(g) Developing virus free plants and synthetic and expression; Important protein databases
seeds. available for the public on the internet like
PDB (Protein Data Bank), PIR (Protein
(h) Biodegradable plastics (concept of PHB). Identification Resources).

309
 PAPER II presterilized petri-dishes under a LAF. To
prepare slants the media is poured into several
PRACTICAL WORK – 15 marks
test tubes, plugged and kept in a tilted position (at
Candidates are required to complete the following an angle of 45o) until it sets.
experiments.
6. Inoculation and incubation of Lactobacillus on the
1. Paper Chromatography – separation of culture medium in the Petri plate.
photosynthetic pigments Use of inoculation loop or inoculation needle for
Take any leaf. Extract chlorophyll in 80% the purpose.
acetone. Take a strip of paper or prepare a thin 7. Identification of bacteria by Gram +ve and Gram
layer of silica gel on a slide. Load chlorophyll –ve (from curd /saliva and/or soil solution)
extract at one end of the paper/gel. Keep paper (i) Prepare a bacterial smear on a slide (ii) Stain
or gel in the rising medium in test tube or jar for with crystal violet stain. (iii) Rinse with water.
about 30 minutes. The rising medium should have (iv) Add a few drops of iodine solution. (v) Add
methanol/ acetic acid, n-butanol or benzene. The few drops of 90 % ethanol (vi) Counterstain with
rising fluid should always be at the bottom below safranin solution (vii) Observe the red and blue
the point of loading of chlorophylls. After 30 colonies under the microscope
minutes, three spots: yellow, bluish green and 8. Action of enzymes on starch under: (a) variable
light green will be observed corresponding to temperature (b) variable substrate concentration –
carotenes, chlorophyll A & chlorophyll B. plotting of K m value by graph
2. Preparation of buffers – phosphate, acetate and (i) Soluble starch solution (0.5% - 1%) to be
borate buffers prepared. Test with iodine. Collect saliva,
This experiment should be done to make the dilute 1: 5, add 1 ml of saliva to 10 ml of
basics clear to the students. Basic calculation for starch solution. Incubate for 15 minutes.
buffer preparation should be known. The Again test for presence of starch with iodine.
approach should be to utilize easily available Also test for the presence of reducing sugars
chemicals at reasonable costs. Phosphate, borate in solution. Repeat the same process at the
and acetate buffers can give the range of pH 4 - variable volumes of starch
pH 9.2 (ii) To study the effect of variable temperature on
3. Preparation of culture media the activity of the enzyme salivary amylase.
(i) Bacterial culture Media - Luria Bertani (L.B.) 9. Isolation of DNA from plants
media - Peptone/ Tryptone, yeast extract and Take half a ripe and peeled banana into a beaker
NaCl. (Nutrient broth / Nutrient Agar). and add 50 ml of extraction fluid (1.5gm table salt
+10 ml liquid detergent +90 ml distilled water).
(ii) Plant Tissue culture medium (Sugars +
Place the beaker in a water bath set at 60 degrees
Coconut milk + Agar Agar). C for 15 minutes. Stir gently with a glass rod.
4. Sterilization of culture medium and other Filter 5ml of cooled content into a clean test tube
materials. and add 5ml of cold 90% ethanol. DNA molecules
separate out and appear as white fibres. [DNA
(i) Dry Physical method – heat or radiation. can also be extracted from pea seeds and soaked
(ii) Wet Physical methods – steam sterilization. wheat grains]

(iii) Chemical Sterilization/ Surface sterilization 10. DNA estimation by colorimeter by DPA method.
Disinfection with 70% alcohol and Sodium 11. Protein estimation by colour reaction – Bradford
hypochlorite solution carbolic acid test.
5. Preparation of various forms of culture media – Bradford’s Assay is a Dye binding assay based on
Petri plate, slant and suspension. the differential change of colour of a dye in
Luria Bertani (L.B) media to be prepared, response to various concentrations of proteins.
autoclaved and cooled to 60 degrees C. To Bradford’s assay can be performed for qualitative
prepare nutrient plates the media is poured into

310
 as well as quantitative assessment of proteins in a A list of suggested projects is as follows:
sample. 1. Effluent analysis.
Dilute 1 volume of Bradford’s dye with 4 volumes 2. A study of the technological details of malt
preparation.
of distilled water. Filter the dye through Whatman
filter paper and store at room temperature in a 3. A study of the technological details of the
brewing industry.
brown glass bottle. Take different aliquots of
standard Bovine Serum Albumin (BSA solution), 4. A study of the organisation of a fermenter.
for example (0.2, 0.4, 0.6, 0.8 and 1.0 ml) in 5. Technological analysis of the process of drug
different test tubes Make up the volume to 1ml development, drug designing and drug targeting.
with distilled water. To each tube add 2ml of 6. A study of the technological details of vaccine
Bradford’s dye. Extent of colour development can development.
be made by rough estimate using + signs to show 7. Diagnosis of diseases by modern techniques like
the concentration of protein in the sample. ELISA, RIA and Antibody targeting.
Alternatively, OD can be read using colorimeter 8. DNA finger-printing.
or spectrophotometer. Take the unknown sample 9. DNA foot-printing.
to be estimated and perform the experiment. 10. Microbiological contaminants in food and food
Similarly read the OD and note the corresponding products.
concentration of protein in it using the graph. 11. Isolation of microbes from air, water and soil.
12. Cell viability test by Evan’s blue dye. 12. Methods of identifying microbes (various
staining techniques and biochemical reactions).
13. Isolation of milk protein – wet weight and dry
13. Tissue Culture and its applications.
weight.
14. Stem Cell Technology
Milk proteins are isolated by adding 0.4 N HCl 15. Nanotechnology
into the milk sample. Casein start coagulating at
16. Bioinformatics
its isoelectric point (i.e. at pH 4.6). The
17. Genetic Engineering
precipitate is filtered and weighed to quantify the
protein present. 18. Cloning
19. Instrumentation in biotechnology
14. Chromatography to find adulteration in spices by
20. Forensic Biotechnology
using mixer of turmeric and metanil yellow.
21. Ethical, Legal and Social Issues (ELSI) related to
15. Demonstration of cell counting by Biotechnology/ GMOs
haemocytometer by using diluted blood. 22. Biopiracy- Case Studies
16. Experiment to show the process of saponification.
Practical File – 5 Marks
PROJECT WORK AND PRACTICAL FILE The Visiting Examiner is required to assess students
on the basis of the practical file maintained by them
– 15 Marks during the academic year.
Project Work – 10 Marks Suggested Evaluation Criteria for Project Work:
Format of the Project:
The Project Work is to be assessed by a Visiting
Examiner appointed locally and approved by the – Content
Council. – Introduction
– Presentation (graphs, tables, charts, newspaper
Candidates are to creatively execute one cuttings, handmade diagrams, photographs,
project / assignment on an aspect of Biotechnology. statistical analysis if relevant)
– Conclusion/ Summary
Teachers may assign or students may choose any one – Bibliography
project of their choice. The report should be kept
simple, but neat and elegant. No extra credit shall be Projects should be handwritten by the candidate.
given for type-written material/decorative cover, etc. Written pages should not exceed 15-20 pages.
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