BIOTECHNOLOGY (878)

Note: The Syllabus for this Subject has not been changed.
CLASS XII

There will be two papers in the subject:

Paper I: Theory……………. 3 hours ... 70 marks Paper II: Practical…………. 3 hours ... 15 marks
Project Work………. … 10 marks
Practical File………… … 5 marks

PAPER I: THEORY- 70 Marks

1. Molecular Biology (a) Concept of transcriptional unit,
promoter, structural and terminator
(i) Nucleic acids and their estimation: an
region; concept of split gene - intron and
understanding of nucleic acids, their
exon; monocistronic and polycistronic
biochemical structure.
RNA, hnRNA;
DNA as the genetic material (Hershey and
(b) Transcription – explanation of the
Chase experiment).
complete process including enzymes
DNA (B-DNA)– physical and chemical involved in the process; Post-
structure; definition, double helical model of transcriptional changes and their
DNA, (Watson and Crick’s); Nucleotide and significance in eukaryotes –
nucleoside; Chargaff’s Law, method of polyadenylation, capping and RNA
replication of DNA, various replicative splicing;
enzymes in both prokaryotic and eukaryotic
(c) Concept of reverse transcription;
organisms, example topoisomerases,
helicase, SSBs polymerases, primases, (d) Genetic code – properties of genetic
ligases. Concept of semi conservative (with code, start and stop codons, anticodons.
respect to Messelson and Stahl experiment
(e) The translation of RNA to protein –
and Taylor et.al experiment on Vicia faba
complete mechanism of chain initiation,
using radiolabelled thymidine) and semi-
elongation and termination, the role of
discontinuous replication, (leading and
tRNA, mRNA and rRNA in protein
lagging strands), okazaki fragments.
synthesis. (Post translational changes
RNA – definition, various types of RNAs such not included).
as mRNA, tRNA (Clover leaf model with
(iii) Gene regulation in prokaryotes
diagram; brief introduction to L-shaped
model), rRNA their structure and functions. Operon concept – lac operon and trp
operon.
Techniques of nucleic acid estimation –
colorimetry and UV-visible 2. Genetic Engineering
spectrophotometry. (i) Introduction to gene cloning and genetic
(ii) Protein Synthesis: synthesis of different engineering: concept of cloning and vectors.
RNAs, and the complete mechanism of Tools of recombinant DNA technology, types
polypeptide chain formation. of restriction endonucleases and other
Concept of central dogma. enzymes used in gene cloning; techniques
involved in extraction and purification of DNA
From genes to proteins:
from bacterial, plant and animal cells.

1
 Selection of host cells: eukaryotic and (iii) Gene analysis techniques: various techniques
prokaryotic. involved in recombinant DNA technology.
Vectors: Characteristics and types such as DNA probes – definition and use.
plasmids -pBR322, pUC (in pBR322- presence
of two antibiotic resistant genes and in Low resolution mapping techniques: gel
pUCpresence of lac Z gene to be taught), electrophoresis, southern blotting (details of
cosmids, phages (M13 and λ), YACs, BACs (to the technique to be taught), western and
be taught with reference to stability and their northern blotting (a brief idea and their
carrying capacity), animal and plant viruses uses).
(CaMV, retrovirus, SV40 – only names of High resolution techniques: DNA
viruses, no details). sequencing- sequencing by chain
Transfer of recombinants into host cells – termination, automated DNA sequencing.
Site directed mutagenesis.
(a) Vectorless methods - basic concept of
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
finger printing.
(b) Vector-mediated method - Agrobacterium
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 respectively.
resistance). (i) General tools and techniques used in cell
A basic understanding of DNA libraries – culture technology
construction of genomic and cDNA (a) Instruments - centrifuge, LAF hood and
libraries. biosafety cabinets, pH meter, autoclave,
Construction of a recombinant DNA vortex mixer, hot air oven, magnetic
molecule. stirrer, weighing balance, micro
filtration unit, incubator, CO2
(ii) Innovations in Biotechnology: produced by incubator, inverted microscope,
using modern biotechnological tools, (select bioreactor (diagram, its components
examples of products already available) and their function)-stirred tank and
sparged type (brief idea only), use of T
(a) Plants: Production of Flavr Savr flasks to propagate animal cells.
tomatoes, Bt-crops and Golden rice.
Only uses of the above instruments to
(b) Healthcare: Production of recombinant
be studied.
hepatitis-B vaccine, Humulin, interferon
and edible vaccines. (b) Sterilization techniques for culture
(c) Animal: Dolly the cloned sheep, Sources room, apparatus, transfer area, media,
and characteristics of stem cells and vitamins, and living material;
their applications. (c) Cryopreservation (need and steps).

(d) Environmental biotechnology: (d) Cell counting (direct counting by

bioremediation using oil-eating bacteria haemocytometer), cell viability by
as an example. Evan’s blue stain and cell sorting
(FACS only)
(e) Industrial biotechnology: applications
of industrial enzymes – rennet,
subtilisin, amylase, papain.
2
 (e) Types of media (synthetic /defined, (h) Biodegradable plastics (concept of PHB).
semi-synthetic/differential,
(iv) Animal cell culture and its application.
complex/natural)
Primary cell culture with mechanical and
Preparation of media: microbial media- enzymatic disaggregation and its drawbacks;
LB agar and LB broth; Plant media-MS Types of cell-lines: finite, continuous,
and White’s media; Animal media- adherent and suspension; scale up-mono
RPMI, DMEM and FBS - brief idea layer by Roller bottle, application of animal
cell culture-tissue, hybridoma technology,
only. (includes inorganic and organic tissue engineering (definition only).
macronutrients and micronutrients,
4. Bioinformatics
antibiotics, growth regulators for
(i) Introduction to bioinformatics; global
plants: auxins and cytokinins).
bioinformatics databases and data retrieval
Importance of pH and solidifying agents. tools; genomics, different types of sequences,
types of sequence analysis.
(ii) Microbial culture and its application. Introduction to bioinformatics: definition and
Fermentation process and growth kinetics- need.
batch culture, fed batch culture, continuous An introduction to global bioinformatics
culture (with the help of graphs only): databases (nucleotide and protein
Definition of turbidostat and chemostat: databases). Information sources such as
Products and application-SCP (definition EMBL, NCBI, DDBJ, SWISSPROT,
and use), industrial enzyme-subtilisin (source GenBank, GENSCAN.
and its use). Data retrieval tools- ENTREZ, Taxonomy
Browser.
(iii) Plant tissue culture and its application.
(ii) Genomics: Definition, introduction, tools used
Isolation of single cell by mechanical and in Genomics and its applications.
enzymatic methods, synchronisation of cell Definition of genomics. Types of genomics-
culture by chemical methods like starvation, structural and functional. Basic criteria in
inhibition and mitotic arrest. selecting the organism for its genome
Cellular totipotency-definition of cellular sequencing. Different types of sequences –
differentiation, de-differentiation, re- cDNA, genomic DNA, ESTs (Expressed
differentiation. Application of plant cell Sequence Tags) and STSs (Sequence Tagged
culture technology (methodology not Sites) and the different softwares (example
required, only brief idea needed): gene scan).
(a) Haploid production-androgenesis and Types of sequence analysis by using BLAST
and FASTA –global, local, pair wise and
gynogenesis and their significance.
multiple.
(b) Triploid production-understanding and
Human Genome Project - its objectives, the
need for triploid production and its countries involved, its achievements and
application (seedless crops). significance.
(c) In-vitro pollination- concept and its DNA microarray technology – definition and
application. application only.
(d) Zygotic embryo culture- concept and its Concept of Single Nucleotide Polymorphisms
application, Embryo rescue (brief idea (SNPs).
only). (iii) Proteomics: definition, introduction and
(e) Somatic hybridisation-protoplast fusion databases.
(Pomato). Types of Proteomics – structural, functional
(f) Micropropagation and its significance. and expression; Important protein databases
(g) Developing virus free plants and synthetic available for the public on the internet like
seeds. PDB (Protein Data Bank), PIR (Protein
Identification Resources).
3
 PAPER II test tubes, plugged and kept in a tilted position
(at an angle of 45o) until it sets.
PRACTICAL WORK – 15 marks
6. Inoculation and incubation of Lactobacillus on
Candidates are required to complete the following the culture medium in the Petri plate.
experiments.
Use of inoculation loop or inoculation needle for
1. Paper Chromatography – separation of the purpose.
photosynthetic pigments
7. Identification of bacteria by Gram +ve and Gram
Take any leaf. Extract chlorophyll in 80% –ve (from curd /saliva and/or soil solution)
acetone. Take a strip of paper or prepare a thin
layer of silica gel on a slide. Load chlorophyll (i) Prepare a bacterial smear on a slide (ii) Stain
extract at one end of the paper/gel. Keep paper with crystal violet stain. (iii) Rinse with water.
or gel in the rising medium in test tube or jar for (iv) Add a few drops of iodine solution. (v) Add
about 30 minutes. The rising medium should few drops of 90 % ethanol (vi) Counterstain with
have methanol/ acetic acid, n-butanol or safranin solution (vii) Observe the red and blue
benzene. The rising fluid should always be at the colonies under the microscope
bottom below the point of loading of 8. Action of enzymes on starch under: (a) variable
chlorophylls. After 30 minutes, three spots: temperature (b) variable substrate concentration –
yellow, bluish green and light green will be plotting of Km value by graph
observed corresponding to carotenes, (i) Soluble starch solution (0.5% - 1%) to be
chlorophyll A & chlorophyll B. prepared. Test with iodine. Collect saliva,
2. Preparation of buffers – phosphate, acetate and dilute 1: 5, add 1 ml of saliva to 10 ml of
borate buffers starch solution. Incubate for 15 minutes.
This experiment should be done to make the Again test for presence of starch with iodine.
basics clear to the students. Basic calculation for Also test for the presence of reducing sugars
buffer preparation should be known. The in solution. Repeat the same process at the
approach should be to utilize easily available variable volumes of starch
chemicals at reasonable costs. Phosphate, borate (ii) To study the effect of variable temperature on
and acetate buffers can give the range of pH 4 - the activity of the enzyme salivary amylase.
pH 9.2 9. Isolation of DNA from plants
3. Preparation of culture media Take half a ripe and peeled banana into a beaker
(i) Bacterial culture Media - Luria Bertani and add 50 ml of extraction fluid (1.5gm table
(L.B.) media - Peptone/ Tryptone, yeast salt +10 ml liquid detergent +90 ml distilled
extract and NaCl. (Nutrient broth / Nutrient water). Place the beaker in a water bath set at 60
Agar). degrees C for 15 minutes. Stir gently with a glass
(ii) Plant Tissue culture medium (Sugars + rod. Filter 5ml of cooled content into a clean test
Coconut milk + Agar Agar). tube and add 5ml of cold 90% ethanol. DNA
molecules separate out and appear as white
4. Sterilization of culture medium and other fibres. [DNA can also be extracted from pea
materials. seeds and soaked wheat grains]
(i) Dry Physical method – heat or radiation. 10. DNA estimation by colorimeter by DPA method.
(ii) Wet Physical methods – steam sterilization. 11. Protein estimation by colour reaction – Bradford
(iii) Chemical Sterilization/ Surface sterilization test.
Disinfection with 70% alcohol and Sodium Bradford’s Assay is a Dye binding assay based
hypochlorite solution carbolic acid on the differential change of colour of a dye in
5. Preparation of various forms of culture media – response to various concentrations of proteins.
Petri plate, slant and suspension. Bradford’s assay can be performed for
Luria Bertani (L.B) media to be prepared, qualitative as well as quantitative assessment of
autoclaved and cooled to 60 degrees C. To proteins in a sample.
prepare nutrient plates the media is poured into Dilute 1 volume of Bradford’s dye with 4 volumes
presterilized petri-dishes under a LAF. To of distilled water. Filter the dye through
prepare slants the media is poured into several Whatman filter paper and store at room
4
 temperature in a brown glass bottle. Take A list of suggested projects is as follows:
different aliquots of standard Bovine Serum 1. Effluent analysis.
Albumin (BSA solution), for example (0.2, 0.4, 2. A study of the technological details of malt
0.6, 0.8 and 1.0 ml) in different test tubes Make preparation.
up the volume to 1ml with distilled water. To 3. A study of the technological details of the
each tube add 2ml of Bradford’s dye. Extent of brewing industry.
colour development can be made by rough 4. A study of the organisation of a fermenter.
estimate using + signs to show the concentration 5. Technological analysis of the process of drug
of protein in the sample. Alternatively, OD can development, drug designing and drug targeting.
be read using colorimeter or spectrophotometer. 6. A study of the technological details of vaccine
Take the unknown sample to be estimated and development.
perform the experiment. Similarly read the OD
7. Diagnosis of diseases by modern techniques like
and note the corresponding concentration of
ELISA, RIA and Antibody targeting.
protein in it using the graph.
8. DNA finger-printing.
12. Cell viability test by Evan’s blue dye. 9. DNA foot-printing.
13. Isolation of milk protein – wet weight and dry 10. Microbiological contaminants in food and food
weight. products.
11. Isolation of microbes from air, water and soil.
Milk proteins are isolated by adding 0.4 N HCl
12. Methods of identifying microbes (various
into the milk sample. Casein start coagulating at
staining techniques and biochemical reactions).
its isoelectric point (i.e. at pH 4.6). The
precipitate is filtered and weighed to quantify the 13. Tissue Culture and its applications.
protein present. 14. Stem Cell Technology
15. Nanotechnology
14. Chromatography to find adulteration in spices by 16. Bioinformatics
using mixer of turmeric and metanil yellow.
17. Genetic Engineering
15. Demonstration of cell counting by 18. Cloning
haemocytometer by using diluted blood. 19. Instrumentation in biotechnology
16. Experiment to show the process of 20. Forensic Biotechnology
saponification. 21. Ethical, Legal and Social Issues (ELSI) related
to Biotechnology/ GMOs
PROJECT WORK AND PRACTICAL FILE 22. Biopiracy- Case Studies

– 15 Marks Practical File – 5 Marks

Project Work – 10 Marks The Visiting Examiner is required to assess students
on the basis of the practical file maintained by them
The Project Work is to be assessed by a Visiting during the academic year.
Examiner appointed locally and approved by the
Suggested Evaluation Criteria for Project Work:
Council.
Format of the Project:
Candidates are to creatively execute one
project / assignment on an aspect of Biotechnology. – Content
– Introduction
Teachers may assign or students may choose any one – Presentation (graphs, tables, charts, newspaper
project of their choice. The report should be kept cuttings, diagrams, photographs, statistical
simple, but neat and elegant. analysis if relevant)
– Conclusion/ Summary
– Bibliography

5
 LIST OF EQUIPMENT FOR BIOTECHNOLOGY PRACTICALS FOR CLASSES XI & XII
1. Table-top Centrifuge 12. Incubator
2. Vortex - Mixer 13. Magnetic stirrer with hot plate
3. Thermostatic water-bath 14. Laminar flow cabinet
4. Spectrophotometer (UV visible range)/ 15. Weighing Balance (Electrical)
Colorimeter 16. Hot plate
5. Refrigerator 17. Binocular Microscope
6. Lactometer 18. Haemocytometer
7. pH meter 19. Colony counter
8. Hot air oven 20. Antiserum
9. Autoclave 21. Electrophoresis chamber
10. Desiccators 22. Micropipettes
11. Micro-filtration unit
LIST OF ABBREVIATIONS TO BE STUDIED
1. BAC: Bacterial Artificial Chromosomes 21. NCBI: National Centre for Biotechnology
2. BLAST: Basic Local Alignment Search Tool Information
3. CTAB: Cetyl Trimethyl Ammonium Bromide 22. NHGRI: National Human Genome Research
4. DBM: Diazo–benzyl oxy–methyl paper Institute
5. DDBJ: DNA Database/ Data Bank of Japan 23. PAGE: Polyacrylamide Gel Electrophoresis
6. ddNTP: Dideoxy Nucleoside triphosphate 24. PCR: Polymerization Chain Reaction
7. DMEM: Dulbecco Modified Eagle Medium 25. PDB: Protein Database/ Data Bank
8. EBI: European Bioinformatics Institute 26. PHB: Poly 3–Hydroxyl Butyrate
9. EMBL: European Molecular Biology Laboratory 27. PIR: Protein Information Resource
10. EST: Expressed Sequence Tag 28. RFLP: Restriction Fragment Length
11. FACS: Fluorescence Activated Cell Sorting Polymorphism
12. FASTA: Fast All 29. RNA: Ribonucleic acid
13. FBS: Foetal Bovine Serum 30. RPMI medium: Roswell Park Memorial Institute
14. HEPA: High Energy Particulate Air medium
15. HGP: Human Genome Project 31. SCP: Single Cell Protein
16. IBPGR: International Board of Plant Genetic 32. SDS – PAGE: Sodium Dodecyl Sulphate–
Resources Polyacrylamide Gel Electrophoresis
17. ICGEB: International Centre for Genetic 33. SNP: Single Nucleotide Polymorphism
Engineering and Biotechnology 34. SSBs: Single Stranded Binding Proteins
18. IFN: Interferon 35. STS: Sequence Tagged Site
19. LB medium: Luria and Bertani Medium 36. VNTR: Variable Number of Tandem Repeats
20. MS medium: Murashige and Skoog medium 37. YAC: Yeast Artificial Chromosome