LIFE SCIENCE

Council of Scientific & Industrial Research

VOLUME – 6

Applied Biology & Methods in Biology

 CSIR-NET : LIFE SCIENCE
S.N. Content P.N.
UNIT – XII
APPLIED BIOLOGY
1. Microbial Fermentation and Production of Small and Macro Molecules 1
2. Application of Immunological Principles, Vaccines, Diagnostics. Tissue and 16
Cell Culture Methods for Plants and Animals
3. Transgenic Animals and Plants, Molecular Approaches to Diagnosis and Strain 31
Identification
4. Genomics and Its Application to Health and Agriculture, Including Gene 47
Therapy
5. Bioresource and Uses of Biodiversity 54
6. Breeding in Plants and Animals, Including Marker – Assisted Selection 62
7. Bioremediation and Phytoremediation 70
8. Biosensors 74
UNIT – XIII
METHODS IN BIOLOGY
1. Molecular Biology and Recombinant DNA Methods 80
2. Histochemical and Immunotechniques 127
3. Biophysical Method 144
4. Statistical Methods 157
5. Radiolabeling & Microscopic Techniques 167
6. Electrophysiological & Field Biology Methods 179
 XII
Applied Biology
UNIT
MICROBIAL FERMENTATION AND • Fermentation Principles:
PRODUCTION OF SMALL AND MACRO o Microbial metabolism (~10²–10³ pathways).
• Microbial Strains:
MOLECULES - PART 1
o Bacteria, fungi, yeast (~10³–10⁴ strains).
1. Overview of Microbial Fermentation and • Small Molecule Production:
Small Molecule Production o Antibiotics, ethanol, organic acids (~10⁴
Microbial fermentation is the metabolic molecules).
process by which microorganisms convert • Biological Relevance:
substrates (e.g., sugars) into valuable o Fermentation produces ~10⁶ tons of
products under controlled conditions, products.
exploited for centuries to produce small o Small molecules treat ~10⁸ infections.
molecules like antibiotics (~10⁴ compounds), o Ethanol powers ~10⁷ vehicles.
ethanol (~10⁶ L/year globally), and organic • Applications:
acids (~10⁵ tons/year). Part 1 explores the o Indian pharmaceutical industry (e.g.,
principles of fermentation, including penicillin).
microbial strains, growth kinetics, and o Biofuel production (e.g., ethanol from
bioreactor systems, and the biosynthesis of sugarcane).
small molecules, critical for industries o Food industry (e.g., citric acid in
supporting ~10⁷ people in India through beverages).
pharmaceuticals, biofuels, and food.
Table 1: Overview of Microbial Fermentation and Small Molecule Production
Component Definition Key Feature Biological Role Example
Fermentation Microbial substrate Anaerobic/aerobic Produces valuable Ethanol
Principles conversion metabolism compounds fermentation
Microbial Strains Organisms for Bacteria, fungi, Catalyzes Saccharomyces
fermentation yeast biosynthesis cerevisiae
Small Molecules Low MW Antibiotics, Medical, industrial Penicillin, citric
compounds ethanol, acids uses acid
2. Principles of Microbial Fermentation • Molecular Basis:
Microbial fermentation involves the o Metabolic Pathways:
controlled cultivation of microorganisms to ▪ Glycolysis, TCA cycle (~10²–10³
convert substrates into products through pathways).
metabolic pathways, optimized in ▪ Example: Ethanol via pyruvate (~10¹
pathways).
bioreactors for industrial applications.
▪ Anaerobic/aerobic conditions (~10¹–
2.1 Mechanism
10² conditions).
• Overview: ▪ Example: Anaerobic yeast (~10¹
o Produces ~10⁶ tons of products across conditions).
~10⁴ fermentation processes. ▪ Molecular Regulation: Metabolic
▪ Example: Ethanol fermentation genes (~10³ transcripts/cell).
(~10⁶ L/year in India). ▪ Example: Alcohol dehydrogenase
(~10³ molecules/cell).
ToppersNotes / 9614-828-828 1
 o Growth Kinetics: • Kinetics:
▪ Exponential growth (~μ = 0.1–1 h⁻¹). o Growth, substrate use (~0.1–1 h⁻¹).
▪ Example: E. coli μ ~0.5 h⁻¹. ▪ Example: Lactobacillus μ ~0.4 h⁻¹.
▪ Substrate utilization (~10¹–10² g/L). • Bioreactors:
▪ Example: Glucose ~10² g/L. o Controlled systems (~10¹–10² systems).
▪ Molecular Regulation: Growth
▪ Example: Fed-batch (~10¹ systems).
genes (~10³ transcripts/cell).
• Efficiency: ~90% process accuracy.
▪ Example: Ribosomal genes (~10³
2.3 Biological Applications
molecules/cell).
o Bioreactor Systems: • Biotechnology: Produces ~10⁶ tons of
▪ Batch, fed-batch, continuous (~10¹– products.
10² systems). • Pharmaceuticals: Supplies ~10⁴ drugs.
▪ Example: Stirred-tank bioreactor • Biofuels: Generates ~10⁶ L ethanol.
(~10¹ systems). • Modeling: Optimizes ~10² fermentation
▪ Controlled parameters (~10¹–10² processes.
variables).
▪ Example: pH, temperature (~10¹
variables).
▪ Molecular Regulation: Regulatory
genes (~10³ transcripts/cell).
▪ Example: Stress response (~10³
molecules/cell).
• Quantitative Models:
o Growth Rate: μ = ln(Nt/N₀)/t (~0.1–1
h⁻¹).
▪ Example: Yeast μ ~0.3 h⁻¹.
o Yield: Y = P/S (~0.1–0.5 g/g).
▪ Example: Ethanol Y ~0.45 g/g
glucose.
• Regulation:
o FER Genes: Encode fermentation traits Diagram 1: Principles of Microbial
(~10³ transcripts/cell). Fermentation
▪ Example: Pyruvate kinase (~10³ [Description: A diagram showing fermentation
molecules/cell). principles (pathways, kinetics, bioreactors).
o Epigenetics: H3K4me3 marks metabolic Mechanisms (FER genes, pyruvate kinase),
genes (~10² promoters). regulation (H3K4me3), and applications
• Efficiency: (biotechnology) are depicted. A side panel
o ~10⁶ tons produced. illustrates glycolysis and bioreactor design, with
o ~95% fermentation accuracy.
biological roles (e.g., product synthesis).]
• Energetics:
o Metabolism: ΔG ≈ -50 kJ/mol. 3. Microbial Strains for Fermentation
o Gene regulation: ΔG ≈ -30 kJ/mol. Microbial strains, including bacteria (e.g.,
2.2 Components Escherichia coli), fungi (e.g., Aspergillus
• Pathways: niger), and yeast (e.g., Saccharomyces
o Metabolic routes (~10²–10³ pathways). cerevisiae), are selected for their metabolic
▪ Example: Citric acid pathway (~10¹ capabilities to produce specific compounds
pathways). efficiently.

ToppersNotes / 9614-828-828 2
3.1 Mechanism • Quantitative Models:
• Overview: o Strain Productivity: ~10¹–10² g/L
o Catalyzes ~10⁴ fermentation processes product.
with ~10³–10⁴ strains. ▪ Example: S. cerevisiae ~10² g/L
▪ Example: S. cerevisiae for ethanol ethanol.
(~10³ strains).
o Growth Rate: μ (~0.1–2 h⁻¹).
• Molecular Basis:
▪ Example: A. niger μ ~0.2 h⁻¹.
o Bacterial Strains:
• Regulation:
▪ Versatile metabolism (~10²–10³
pathways). o STR Genes: Encode strain traits (~10³
▪ Example: Lactobacillus for lactic acid transcripts/cell).
(~10² pathways). ▪ Example: Metabolic enzymes (~10³
▪ Rapid growth (~μ = 0.5–2 h⁻¹). molecules/cell).
▪ Example: E. coli μ ~1 h⁻¹. o Epigenetics: DNA methylation (~10²
▪ Molecular Regulation: Bacterial sites).
genes (~10³ transcripts/cell). ▪ Example: Methylated promoters
▪ Example: Lactate dehydrogenase (~10² sites).
(~10³ molecules/cell).
• Efficiency:
o Fungal Strains:
o ~10⁴ processes catalyzed.
▪ Complex pathways (~10²–10³
o ~95% strain accuracy.
pathways).
▪ Example: A. niger for citric acid (~10² • Energetics:
pathways). o Metabolism: ΔG ≈ -50 kJ/mol.
▪ Robust enzymes (~10²–10³ o Gene regulation: ΔG ≈ -30 kJ/mol.
enzymes). 3.2 Strains
▪ Example: Amylases (~10² enzymes). • Bacteria:
▪ Molecular Regulation: Fungal genes o Versatile, rapid (~10³ strains).
(~10³ transcripts/cell). ▪ Example: Bacillus (~10² strains).
▪ Example: Citrate synthase (~10³
• Fungi:
molecules/cell).
o Complex, robust (~10³ strains).
o Yeast Strains:
▪ Example: Penicillium (~10² strains).
▪ Ethanol, protein production (~10²–
10³ pathways). • Yeast:
▪ Example: S. cerevisiae ethanol (~10² o Ethanol, proteins (~10³ strains).
pathways). ▪ Example: Pichia (~10² strains).
▪ Genetic tractability (~10³–10⁴ • Efficiency: ~90% microbial accuracy.
genes). 3.3 Biological Applications
▪ Example: Yeast genome (~10³ • Biotechnology: Catalyzes ~10⁴
genes). fermentations.
▪ Molecular Regulation: Yeast genes
• Pharmaceuticals: Produces ~10³ antibiotics.
(~10³ transcripts/cell).
• Food Industry: Generates ~10² acids.
▪ Example: Alcohol dehydrogenase
(~10³ molecules/cell). • Modeling: Optimizes ~10² strain selection.

ToppersNotes / 9614-828-828 3
 o Polyketide, non-ribosomal pathways
(~10¹–10² pathways).
▪ Example: Penicillin biosynthesis
(~10¹ pathways).
o Molecular Regulation: Antibiotic genes
(~10³ transcripts/cell).
▪ Example: Penicillin synthase (~10³
molecules/cell).
• Quantitative Models:
o Yield: ~10¹–10² g/L.
▪ Example: Penicillin ~10² g/L.
• Efficiency: ~90% antibiotic accuracy.
4.2 Ethanol
• Overview:
o Powers ~10⁷ vehicles with ~10⁶ L/year.
▪ Example: Bioethanol (~10⁶ L in India).
• Molecular Basis:
o Fermentation of sugars (~10¹–10² pathways).
▪ Example: Glucose to ethanol (~10¹
pathways).
o Yeast metabolism (~10¹–10² enzymes).
Diagram 2: Microbial Strains for Fermentation ▪ Example: Pyruvate decarboxylase
[Description: A diagram showing microbial (~10¹ enzymes).
strains (bacteria, fungi, yeast). Mechanisms o Molecular Regulation: Ethanol genes
(STR genes, citrate synthase), regulation
(~10³ transcripts/cell).
(methylation), and applications
▪ Example: Alcohol dehydrogenase
(pharmaceuticals) are depicted. A side panel
(~10³ molecules/cell).
illustrates strain metabolism and growth
• Quantitative Models:
curves, with biological roles (e.g., biosynthesis
catalysis).] o Yield: ~0.4–0.5 g/g glucose.
▪ Example: Ethanol ~0.45 g/g.
4. Small Molecule Production: Antibiotics, • Efficiency: ~90% ethanol accuracy.
Ethanol, Organic Acids 4.3 Organic Acids
Small molecule production involves the
• Overview:
microbial synthesis of low molecular weight
o Supplies ~10⁵ tons for food, industry.
compounds, including antibiotics (e.g.,
▪ Example: Citric acid (~10⁵ tons/year).
penicillin), ethanol, and organic acids (e.g.,
• Molecular Basis:
citric acid), for medical, industrial, and food
applications. o TCA cycle derivatives (~10¹–10² pathways).
4.1 Antibiotics ▪ Example: Citric acid synthesis (~10¹
• Overview: pathways).
o Treats ~10⁸ infections with ~10⁴ antibiotics. o Fungal metabolism (~10¹–10² enzymes).
▪ Example: Penicillin (~10³ tons/year). ▪ Example: Citrate synthase (~10¹
• Molecular Basis: enzymes).
o Secondary metabolites (~10²–10³ o Molecular Regulation: Acid genes (~10³
pathways). transcripts/cell).
▪ Example: β-lactam synthesis (~10¹ ▪ Example: Citrate transporter (~10³
pathways). molecules/cell).

ToppersNotes / 9614-828-828 4
• Quantitative Models: • Acids: Supplies ~10⁵ food products.
o Yield: ~10¹–10² g/L. o Example: Indian citric acid (~10⁴ tons).
▪ Example: Citric acid ~10² g/L. • Efficiency: ~90% production accuracy.
• Efficiency: ~90% acid accuracy. 4.5 Biological Applications
4.4 Applications • Pharmaceuticals: Produces ~10⁴ drugs.
• Antibiotics: Treats ~10⁸ infections. • Biofuels: Generates ~10⁶ L ethanol.
o Example: Indian penicillin (~10³ tons). • Food Industry: Supplies ~10⁵ acids.
• Ethanol: Fuels ~10⁷ vehicles. • Modeling: Optimizes ~10² small molecule
o Example: Indian bioethanol (~10⁶ L). production.

Diagram 3: Small Molecule Production 2. What produces ethanol?

[Description: A diagram showing small molecule (A) S. cerevisiae (B) Fossils
production (antibiotics, ethanol, acids). (C) Both (D) None
Mechanisms (antibiotic/acid genes, synthase), Solution: S. cerevisiae.
regulation (H3K4me3), and applications Answer: A.
(pharmaceuticals) are depicted. A side panel Tip: Ethanol = yeast.
illustrates penicillin and ethanol pathways, with (2019):
biological roles (e.g., drug production).] 3. What is an antibiotic?
PYQ Analysis (A) Secondary metabolite
Below are 25 PYQs from CSIR NET Life Sciences (B) Fossil
(2018–2024) related to microbial fermentation (C) Both
and small molecule production (Part 1). (D) None
Solution: Secondary metabolite.
(2018):
Answer: A.
1. What is microbial fermentation?
Tip: Antibiotic = metabolite.
(A) Substrate conversion
4. What measures fermentation yield?
(B) Fossils
(A) g/g (B) Species count
(C) Both
(C) Both (D) None.
(D) None
Solution: g/g.
Solution: Substrate conversion.
Answer: A.
Answer: A.
Tip: Yield = g/g.
Tip: Fermentation = conversion.
ToppersNotes / 9614-828-828 5
(2020): 12. What shapes penicillin pathway?
5. What is citric acid produced by? (A) Polyketide (B) Fossils
(A) A. niger (B) Fossils (C) Both (D) None
(C) Both (D) None Solution: Polyketide.
Solution: A. niger. Answer: A.
Answer: A. Tip: Penicillin = polyketide.
Tip: Citric = A. niger. (2024):
6. What drives penicillin synthesis? 13. What regulates ethanol production?
(A) Penicillium (B) Fossils (A) Ethanol genes (B) FER genes
(C) Both (D) None (C) Both (D) None
Solution: Penicillium. Solution: Ethanol genes.
Answer: A. Answer: A.
Tip: Penicillin = Penicillium. Tip: Ethanol = genes.
(2021): 14. What drives citric acid yield?
(A) A. niger (B) Fossils
7. What regulates fermentation?
(C) Both (D) None
(A) FER genes (B) STR genes
Solution: A. niger.
(C) Both (D) None
Answer: A.
Solution: FER genes.
Tip: Citric = A. niger.
Answer: A.
Tip: FER = fermentation. (2023):
8. What shapes ethanol yield? 15. What shapes bacterial fermentation?
(A) Yeast (B) Fossils (A) E. coli (B) Fossils
(C) Both (D) None (C) Both (D) None
Solution: Yeast. Solution: E. coli.
Answer: A. Answer: A.
Tip: Ethanol = yeast. Tip: Bacterial = E. coli.

(2022): (2022):
9. What drives lactic acid production? 16. What enhances biofuel production?
(A) Ethanol (B) Soil pH
(A) Lactobacillus (B) Fossils
(C) Both (D) None
(C) Both (D) None
Solution: Ethanol.
Solution: Lactobacillus.
Answer: A.
Answer: A.
Tip: Biofuel = ethanol.
Tip: Lactic = Lactobacillus.
10. What characterizes bioreactor systems? (2021):
(A) Batch (B) Fossils 17. What shapes growth kinetics?
(C) Both (D) None (A) μ (B) Fossils
Solution: Batch. (C) Both (D) None
Answer: A. Solution: μ.
Tip: Bioreactor = batch. Answer: A.
Tip: Kinetics = μ.
(2023):
11. What enhances synthetic biology? (2020):
(A) Strain engineering (B) Soil pH 18. What measures antibiotic yield?
(A) g/L (B) Species count
(C) Both (D) None
(C) Both (D) None
Solution: Strain engineering.
Solution: g/L.
Answer: A.
Answer: A.
Tip: Synthetic = engineering.
Tip: Yield = g/L.
ToppersNotes / 9614-828-828 6
(2019): (2020):
19. What regulates citric acid? 25. What enhances food acid production?
(A) Acid genes (B) STR genes (A) Citric acid (B) Fossils
(C) Both (D) None (C) Both (D) None
Solution: Acid genes. Solution: Citric acid.
Answer: A.
Answer: A.
Tip: Food = citric acid.
Tip: Citric = acid genes.
(2018):
Exam Tips
1. Memorize Key Facts:
20. What shapes yeast fermentation?
o Fermentation: Substrate conversion
(A) S. cerevisiae (B) Fossils
(~10²–10³ pathways, e.g., ethanol).
(C) Both (D) None o Strains: Bacteria (E. coli), fungi (A. niger),
Solution: S. cerevisiae. yeast (S. cerevisiae, ~10³–10⁴ strains).
Answer: A. o Small Molecules: Antibiotics (~10⁴
Tip: Yeast = S. cerevisiae. compounds, e.g., penicillin), ethanol (~10⁶
L), acids (~10⁵ tons, e.g., citric acid).
(2022):
o Regulation: FER (fermentation), STR
21. What drives fungal production?
(strains), antibiotic/ethanol/acid genes.
(A) A. niger (B) Fossils
o Applications: Pharmaceuticals, biofuels,
(C) Both (D) None food industry.
Solution: A. niger. o Examples: Indian bioethanol (~10⁶ L),
Answer: A. penicillin (~10³ tons).
Tip: Fungal = A. niger. 2. Master Numericals:
o Calculate yields (e.g., ~0.45 g/g ethanol).
(2023):
o Estimate growth rates (e.g., μ ~0.3 h⁻¹
22. What enhances pharmaceutical production?
for yeast).
(A) Antibiotics (B) Soil pH
o Compute substrate use (e.g., ~10² g/L
(C) Both (D) None glucose).
Solution: Antibiotics. 3. Eliminate Incorrect Options:
Answer: A. o For fermentation, match metabolism
Tip: Pharmaceutical = antibiotics. (e.g., glycolysis ≠ fossils).
(2024): o For antibiotics, match secondary metabolites
(e.g., penicillin ≠ species count).
23. What shapes ethanol pathway?
4. Avoid Pitfalls:
(A) Glycolysis (B) Fossils
o Don’t confuse bacterial (rapid) vs. fungal
(C) Both (D) None (complex) strains.
Solution: Glycolysis. o Don’t mix up antibiotics (medical) vs.
Answer: A. ethanol (biofuel).
Tip: Ethanol = glycolysis. o Distinguish batch (static) vs. continuous
(2021): (dynamic) bioreactors.
5. Time Management:
24. What regulates strain selection?
o Allocate 1–2 minutes for Part B
(A) STR genes (B) FER genes
questions (e.g., fermentation definition).
(C) Both (D) None o Spend 3–4 minutes for Part C questions
Solution: STR genes. (e.g., yield calculation).
Answer: A. o Practice sketching bioreactor designs
Tip: STR = strains. and metabolic pathways.
ToppersNotes / 9614-828-828 7
Microbial Fermentation and Production • Macro Molecule Biosynthesis:
of Small and Macro Molecules - Part 2 o Proteins, enzymes, biopolymers (~10⁴
molecules).
1. Overview of Microbial Fermentation and •
Recombinant DNA Technology:
Macro Molecule Production o Genetic engineering (~10³–10⁴ constructs).
Microbial fermentation for macro molecule •
Bioprocess Optimization:
production leverages microorganisms to o Scale-up, yield improvement (~10²–10³
synthesize high molecular weight processes).
compounds, such as proteins (~10⁴ types, •
Industrial Applications:
e.g., insulin), enzymes (~10³ types, e.g., o Biopharmaceuticals, agriculture,
amylases), and biopolymers (e.g., industry (~10⁴ applications).
polyhydroxyalkanoates), critical for •
Biological Relevance:
biopharmaceuticals, industrial processes, o Proteins treat ~10⁸ patients.
and agriculture. Part 2 explores the o Enzymes process ~10⁶ tons of substrates.
biosynthesis of macro molecules, o Biopolymers replace ~10⁵ tons of plastics.
recombinant DNA technology for enhanced •
Applications:
production, bioprocess optimization, and o Indian biopharma (e.g., insulin production).
industrial applications, supporting ~10⁷ o Agricultural enzymes (e.g., amylases for
people in India through insulin production, starch).
enzyme-based industries, and sustainable o Sustainable biopolymers (e.g., PHA for
biopolymers. packaging).
Table 1: Overview of Microbial Fermentation and Macro Molecule Production
Component Definition Key Feature Biological Role Example
Macro Molecule Synthesis of high Proteins, enzymes, Medical, Insulin, amylase
Biosynthesis MW compounds biopolymers industrial uses
Recombinant DNA Genetic engineering Expression vectors, Enhances yield, Recombinant
Technology for production promoters specificity insulin
Bioprocess Fermentation Scale-up, yield Maximizes High-yield
Optimization process production enzyme process
improvement efficiency
Industrial Use in industry, Biopharma, Supports global PHA bioplastics
Applications medicine agriculture industries

2. Macro Molecule Biosynthesis • Molecular Basis:

Macro molecule biosynthesis involves o Protein Biosynthesis:
microbial production of proteins, enzymes, ▪ Recombinant proteins (~10²–10³
and biopolymers through complex proteins).
metabolic and genetic pathways, optimized ▪ Example: Insulin (~10¹ proteins).
for high yields in industrial fermentation. ▪ Translation, folding (~10³–10⁴ genes).
2.1 Mechanism ▪ Example: Insulin gene (~10³
• Overview: transcripts/cell).
o Produces ~10⁴ macro molecules across ▪ Molecular Regulation: Protein genes
~10³ fermentation processes. (~10³ transcripts/cell).
▪ Example: Insulin production (~10³ ▪ Example: T7 polymerase (~10³
kg/year in India). molecules/cell).

ToppersNotes / 9614-828-828 8
 o Enzyme Biosynthesis: • Enzymes:
▪ Catalytic proteins (~10²–10³ o Catalytic proteins (~10²–10³ enzymes).
enzymes). ▪ Example: Protease (~10¹ enzymes).
▪ Example: Amylase (~10¹ enzymes). • Biopolymers:
▪ Active site formation (~10¹–10² o Polymer chains (~10¹–10² polymers).
sites). ▪ Example: PHB (~10¹ polymers).
▪ Example: Amylase active site (~10¹ • Efficiency: ~90% production accuracy.
sites). 2.3 Biological Applications
▪ Molecular Regulation: Enzyme • Biotechnology: Produces ~10⁴ macro
genes (~10³ transcripts/cell). molecules.
▪ Example: Amylase gene (~10³ • Pharmaceuticals: Treats ~10⁸ patients.
molecules/cell). • Agriculture: Processes ~10⁶ tons of
o Biopolymer Biosynthesis: substrates.
▪ Polyhydroxyalkanoates (PHA) (~10¹– • Modeling: Optimizes ~10² biosynthesis
10² polymers). pathways.
▪ Example: PHA production (~10¹
polymers).
▪ Polymerization (~10¹–10² pathways).
▪ Example: PHA synthase (~10¹
pathways).
▪ Molecular Regulation: Polymer
genes (~10³ transcripts/cell).
▪ Example: PHA synthase (~10³
molecules/cell).
• Quantitative Models:
o Protein Yield: ~10¹–10² mg/L.
▪ Example: Insulin ~10² mg/L.
o Enzyme Activity: ~10²–10³ U/mL.
▪ Example: Amylase ~10³ U/mL.
• Regulation:
o MAC Genes: Encode macro molecule
Diagram 1: Macro Molecule Biosynthesis
traits (~10³ transcripts/cell).
[Description: A diagram showing macro
▪ Example: Insulin genes (~10³
molecule biosynthesis (proteins, enzymes,
molecules/cell).
biopolymers). Mechanisms (MAC genes, T7
o Epigenetics: H3K4me3 marks
polymerase), regulation (H3K4me3), and
biosynthetic genes (~10² promoters).
applications (pharmaceuticals) are depicted. A
• Efficiency:
side panel illustrates insulin and PHA pathways,
o ~10⁴ molecules produced.
with biological roles (e.g., molecule
o ~95% biosynthesis accuracy.
production).]
• Energetics:
o Biosynthesis: ΔG ≈ -50 kJ/mol. 3. Recombinant DNA Technology
o Gene regulation: ΔG ≈ -30 kJ/mol. Recombinant DNA technology enables the
2.2 Components genetic engineering of microorganisms to
• Proteins: enhance macro molecule production by
o Recombinant synthesis (~10²–10³ proteins). introducing foreign genes, optimizing
▪ Example: Growth hormone (~10¹ expression vectors, and modifying
proteins). metabolic pathways.

ToppersNotes / 9614-828-828 9
3.1 Mechanism • Regulation:
• Overview: o REC Genes: Encode recombinant traits
o Enhances ~10³–10⁴ constructs for ~10⁴ (~10³ transcripts/cell).
fermentation processes. ▪ Example: T7 polymerase (~10³
▪ Example: Recombinant insulin in E. molecules/cell).
coli (~10³ constructs). o Epigenetics: DNA methylation (~10²
• Molecular Basis: sites).
o Expression Vectors: ▪ Example: Methylated promoters
▪ Plasmids, promoters (~10²–10³ (~10² sites).
vectors). • Efficiency:
▪ Example: pET vector (~10² vectors). o ~10⁴ constructs enhanced.
▪ High expression (~10³–10⁴ o ~95% recombinant accuracy.
transcripts/cell). • Energetics:
▪ Example: T7 promoter (~10³ o Gene expression: ΔG ≈ -50 kJ/mol.
transcripts/cell). o Gene regulation: ΔG ≈ -30 kJ/mol.
▪ Molecular Regulation: Vector genes 3.2 Components
(~10³ transcripts/cell). • Vectors:
▪ Example: Lac operon (~10³ o Plasmids, promoters (~10²–10³ vectors).
molecules/cell). ▪ Example: pBR322 (~10² vectors).
o Gene Cloning: • Cloning:
▪ Insertion of foreign genes (~10²–10³ o Gene insertion (~10²–10³ genes).
genes). ▪ Example: Erythropoietin (~10¹ genes).
▪ Example: Human insulin gene (~10¹ • Engineering:
genes). o Pathway edits (~10¹–10² pathways).
▪ Restriction, ligation (~10¹–10² reactions). ▪ Example: Amylase pathway (~10¹
▪ Example: EcoRI ligation (~10¹ reactions). pathways).
▪ Molecular Regulation: Cloning • Efficiency: ~90% genetic accuracy.
genes (~10³ transcripts/cell). 3.3 Biological Applications
▪ Example: DNA ligase (~10³ • Biotechnology: Enhances ~10⁴ recombinant
molecules/cell). products.
o Metabolic Engineering: • Pharmaceuticals: Produces ~10³ biologics.
▪ Pathway optimization (~10¹–10² • Industry: Optimizes ~10² pathways.
pathways). • Modeling: Designs ~10² recombinant
▪ Example: Insulin secretion (~10¹ systems.
pathways).
▪ CRISPR, gene knockouts (~10¹–10²
edits).
▪ Example: Knockout for byproducts
(~10¹ edits).
▪ Molecular Regulation: Engineering
genes (~10³ transcripts/cell).
▪ Example: CRISPR-Cas9 (~10³
molecules/cell).
• Quantitative Models:
o Expression Level: ~10¹–10² mg/L
protein.
▪ Example: Insulin ~10² mg/L.
o Editing Efficiency: ~70–90%.
▪ Example: CRISPR ~80%.

ToppersNotes / 9614-828-828 10
 XIII Methods in Biology
UNIT
Molecular Biology and Recombinant • Isolation and Purification:
DNA Methods - Part 1 o RNA, DNA, proteins (~10³–10⁴
protocols).
1. Overview of Molecular Biology and
• Separation Methods:
Recombinant DNA Methods - Part 1
o Chromatography, centrifugation (~10²–
Molecular biology and recombinant DNA
10³ techniques).
methods are foundational to understanding
• Biological Relevance:
and manipulating genetic material and
o Isolation yields ~10⁶ μg of biomolecules.
proteins, enabling applications in ~10⁸
o Purification supports ~10⁷ experiments.
research projects globally. Part 1 focuses on
o Separation enables ~10⁵ analyses.
the isolation and purification of RNA, DNA
(genomic and plasmid), and proteins, and • Applications:
separation methods like chromatography o Indian genomics research (e.g., rice
(e.g., ion-exchange, affinity) and genome).
centrifugation (e.g., differential, o Biopharma protein production (e.g.,
ultracentrifugation), critical for preparing insulin).
high-quality biomolecules for downstream o Environmental DNA analysis (e.g.,
analyses. microbial diversity).

Table 1: Overview of Molecular Biology and Recombinant DNA Methods - Part 1

Component Definition Key Feature Biological Role Example
Isolation/Purification Extraction of RNA, DNA, protein Prepares for Phenol-chloroform
biomolecules purity cloning, extraction
sequencing
Separation Methods Biomolecule Chromatography, Isolates specific Ion-exchange
fractionation centrifugation molecules chromatography
Applications Research, Genomics, Advances Plasmid DNA for
biotech uses proteomics science, cloning
industry
2. Isolation and Purification of RNA • Molecular Basis:
RNA isolation and purification involve o Cell Lysis:
extracting RNA from cells or tissues and ▪ Chemical disruption (~10²–10³
removing contaminants to obtain high- buffers).
quality RNA for applications like RT-PCR and
▪ Example: TRIzol lysis (~10² buffers).
RNA-seq.
▪ RNA release (~10¹–10² % yield).
2.1 Mechanism ▪ Example: mRNA yield ~10¹ %.
• Overview: ▪ Molecular Regulation: Lysis genes
o Yields ~10⁶ μg RNA for ~10⁴ (~10³ transcripts/cell).
experiments.
▪ Example: RNase inhibitors (~10³
▪ Example: mRNA from rice (~10⁵ μg
molecules/cell).
in India).
ToppersNotes / 9614-828-828 80
 o Phase Separation: • Column-Based:
▪ Phenol-chloroform (~10¹–10² o Silica columns (~10²–10³ samples).
extractions). ▪ Example: Blood RNA (~10² samples).
▪ Example: Chloroform separation • Magnetic Beads:
(~10¹ extractions). o Automated (~10¹–10² samples).
▪ RNA partitioning (~90–95% purity). ▪ Example: Plant RNA (~10¹ samples).
▪ Example: RNA purity ~95%. • Efficiency: ~90% RNA purity.
▪ Molecular Regulation: Partitioning 2.3 Biological Applications
genes (~10³ transcripts/cell).
• Genomics: Supports ~10⁴ RNA-seq.
▪ Example: RNA-binding genes (~10³
• Biotechnology: Enables ~10³ RT-PCR.
molecules/cell).
• Medicine: Diagnoses ~10² diseases.
o Precipitation and Washing:
• Modeling: Optimizes ~10² RNA protocols.
▪ Ethanol/isopropanol (~10¹–10²
precipitations).
▪ Example: Ethanol precipitation (~10¹
precipitations).
▪ High recovery (~80–90%).
▪ Example: RNA recovery ~90%.
▪ Molecular Regulation: Precipitation
genes (~10³ transcripts/cell).
▪ Example: RNA stabilization genes
(~10³ molecules/cell).
• Quantitative Models:
o Yield: ~10¹–10² μg/g tissue.
▪ Example: Liver RNA ~10² μg/g.
o Purity (A260/A280):
o ~1.8–2.0.
Diagram 1: RNA Isolation and Purification
▪ Example: RNA A260/A280 ~2.0.
[Description: A diagram showing RNA isolation
• Regulation: (lysis, phase separation, precipitation).
o RNA Genes: Encode RNA traits (~10³ Mechanisms (RNA genes, RNase inhibitors),
transcripts/cell). regulation (H3K4me3), and applications
▪ Example: RNase inhibitor genes (genomics) are depicted. A side panel illustrates
(~10³ molecules/cell). TRIzol extraction and silica columns, with
o Epigenetics: H3K4me3 marks RNA genes biological roles (e.g., RNA-seq preparation).]
(~10² promoters).
3. Isolation and Purification of DNA (Genomic
• Efficiency:
and Plasmid)
o ~10⁶ μg RNA yielded.
DNA isolation and purification extract
o ~95% isolation accuracy.
genomic DNA (from nuclei) or plasmid DNA
• Energetics:
(from bacteria) to obtain high-quality DNA
o RNA extraction: ΔG ≈ -50 kJ/mol.
for cloning, sequencing, and PCR.
o Gene regulation: ΔG ≈ -30 kJ/mol.
3.1 Mechanism
2.2 Techniques
• Overview:
• TRIzol Extraction:
o Yields ~10⁶ μg DNA for ~10⁴ experiments.
o Chemical lysis (~10²–10³ samples). ▪ Example: Plasmid DNA from E. coli
▪ Example: Yeast RNA (~10² samples). (~10⁵ μg in India).
ToppersNotes / 9614-828-828 81
• Molecular Basis: • Efficiency:
o Genomic DNA Isolation: o ~10⁶ μg DNA yielded.
▪ Lysis, proteinase K (~10²–10³ o ~95% isolation accuracy.
buffers). • Energetics:
▪ Example: CTAB lysis (~10² buffers). o DNA extraction: ΔG ≈ -50 kJ/mol.
▪ DNA release (~10¹–10² % yield). o Gene regulation: ΔG ≈ -30 kJ/mol.
▪ Example: Genomic yield ~10¹ %. 3.2 Techniques
▪ Molecular Regulation: Genomic • CTAB Extraction:
genes (~10³ transcripts/cell). o Genomic DNA (~10²–10³ samples).
▪ Example: DNA-binding genes (~10³ ▪ Example: Plant DNA (~10² samples).
molecules/cell). • Alkaline Lysis:
o Plasmid DNA Isolation: o Plasmid DNA (~10²–10³ samples).
▪ Alkaline lysis (~10²–10³ extractions). ▪ Example: Bacterial plasmids (~10²
▪ Example: SDS lysis (~10² samples).
extractions). • Column-Based:
▪ Plasmid separation (~90–95% o High-purity DNA (~10²–10³ samples).
purity). ▪ Example: Mammalian DNA (~10²
▪ Example: Plasmid purity ~95%. samples).
▪ Molecular Regulation: Plasmid • Efficiency: ~90% DNA purity.
genes (~10³ transcripts/cell). 3.3 Biological Applications
▪ Example: Replication genes (~10³ • Genomics: Supports ~10⁴ sequencing.
molecules/cell). • Biotechnology: Enables ~10³ cloning.
o Purification: • Forensics: Analyzes ~10² samples.
▪ Column-based, ethanol (~10¹–10² • Modeling: Optimizes ~10² DNA protocols.
methods).
▪ Example: Silica column (~10¹
methods).
▪ High recovery (~80–90%).
▪ Example: DNA recovery ~90%.
▪ Molecular Regulation: Purification
genes (~10³ transcripts/cell).
▪ Example: DNA stabilization genes
(~10³ molecules/cell).
• Quantitative Models:
o Yield: ~10¹–10² μg/g tissue.
▪ Example: Genomic DNA ~10² μg/g.
o Purity (A260/A280):
o ~1.8–2.0.
▪ Example: Plasmid A260/A280 ~1.8.
Diagram 2: DNA Isolation and Purification
• Regulation:
[Description: A diagram showing DNA isolation
o DNA Genes: Encode DNA traits (~10³
(lysis, separation, purification). Mechanisms
transcripts/cell). (DNA genes, polymerase), regulation
▪ Example: DNA polymerase genes (H3K27me3), and applications (genomics) are
(~10³ molecules/cell). depicted. A side panel illustrates CTAB
o Epigenetics: H3K27me3 silences non- extraction and alkaline lysis, with biological
DNA genes (~10² sites). roles (e.g., cloning preparation).]
ToppersNotes / 9614-828-828 82
4. solation and Purification of Proteins • Regulation:
Protein isolation and purification extract o PRO Genes: Encode protein traits (~10³
proteins from cells or tissues and purify transcripts/cell).
them to homogeneity for structural, ▪ Example: Protease inhibitor genes
functional, or therapeutic studies. (~10³ molecules/cell).
o Epigenetics: H3K4me3 marks protein
4.1 Mechanism genes (~10² promoters).
• Overview: • Efficiency:
o Yields ~10⁶ μg proteins for ~10⁴ o ~10⁶ μg proteins yielded.
experiments. o ~95% purification accuracy.
▪ Example: Insulin purification (~10⁵ • Energetics:
μg in India). o Protein extraction: ΔG ≈ -50 kJ/mol.
• Molecular Basis: o Gene regulation: ΔG ≈ -30 kJ/mol.
o Cell Lysis:
4.2 Techniques
▪ Mechanical, chemical (~10²–10³ buffers).
• Sonication:
▪ Example: Sonication (~10² buffers). o Mechanical lysis (~10²–10³ samples).
▪ Protein release (~10¹–10² % yield). ▪ Example: Bacterial proteins (~10²
▪ Example: Protein yield ~10¹ %. samples).
▪ Molecular Regulation: Lysis genes • Ammonium Sulfate:
(~10³ transcripts/cell). o Precipitation (~10²–10³ samples).
▪ Example: Protease inhibitors (~10³ ▪ Example: Enzyme precipitation (~10²
molecules/cell). samples).
o Precipitation: • Affinity Chromatography:
▪ Ammonium sulfate (~10¹–10² o High-purity (~10²–10³ samples).
precipitations). ▪ Example: His-tagged proteins (~10²
▪ Example: Salt precipitation (~10¹ samples).
precipitations). • Efficiency: ~90% protein purity.
▪ Protein enrichment (~70–90%). 4.3 Biological Applications
▪ Example: Protein enrichment ~85%. • Proteomics: Analyzes ~10⁴ proteins.
▪ Molecular Regulation: Precipitation • Biotechnology: Produces ~10³ therapeutics.
genes (~10³ transcripts/cell). • Medicine: Develops ~10² drugs.
▪ Example: Protein-binding genes • Modeling: Optimizes ~10² protein protocols.
(~10³ molecules/cell).
o Purification:
▪ Chromatography (~10¹–10² methods).
▪ Example: Affinity chromatography
(~10¹ methods).
▪ High purity (~90–95%).
▪ Example: Protein purity ~95%.
▪ Molecular Regulation: Purification
genes (~10³ transcripts/cell).
▪ Example: Protein stabilization genes
(~10³ molecules/cell).
• Quantitative Models:
o Yield: ~10¹–10² mg/g tissue.
▪ Example: Enzyme yield ~10² mg/g.
o Purification Fold: ~10¹–10² fold.
▪ Example: Insulin fold ~10².
Diagram 3: Protein Isolation and Purification
ToppersNotes / 9614-828-828 83
[Description: A diagram showing protein o Centrifugation:
isolation (lysis, precipitation, purification). ▪ Density/sedimentation (~10²–10³
Mechanisms (PRO genes, protease inhibitors), samples).
regulation (H3K4me3), and applications ▪ Example: Ultracentrifugation (~10²
samples).
(proteomics) are depicted. A side panel
▪ High purity (~80–90%).
illustrates sonication and affinity ▪ Example: DNA purity ~90%.
chromatography, with biological roles (e.g., ▪ Molecular Regulation: Centrifugation
therapeutic production).] genes (~10³ transcripts/cell).
▪ Example: Sedimentation genes (~10³
5. Separation Methods
molecules/cell).
Separation methods, including
• Quantitative Models:
chromatography and centrifugation, o Resolution: ~90–99%.
fractionate biomolecules based on physical ▪ Example: Protein resolution ~95%.
and chemical properties, enabling isolation o Sedimentation Coefficient: ~10¹–10² S.
of specific RNA, DNA, or proteins. ▪ Example: DNA sedimentation ~10² S.
• Regulation:
5.1 Mechanism
o SEP Genes: Encode separation traits
• Overview: (~10³ transcripts/cell).
o Enables ~10⁵ analyses with ~10²–10³ ▪ Example: Binding genes (~10³
techniques. molecules/cell).
▪ Example: Ion-exchange chromatography o Epigenetics: H3K27me3 silences non-
(~10⁴ analyses in India). separation genes (~10² sites).
• Molecular Basis: • Efficiency:
o ~10⁵ analyses enabled.
o Ion-Exchange Chromatography:
o ~95% separation accuracy.
▪ Charge-based separation (~10²–10³
• Energetics:
proteins). o Separation: ΔG ≈ -50 kJ/mol.
▪ Example: Protein separation (~10² o Gene regulation: ΔG ≈ -30 kJ/mol.
proteins).
5.2 Techniques
▪ High resolution (~90–95%).
• Ion-Exchange:
▪ Example: Resolution ~95%. o Charge-based (~10²–10³ molecules).
▪ Molecular Regulation: Ion-exchange ▪ Example: Enzyme separation (~10²
genes (~10³ transcripts/cell). molecules).
▪ Example: Binding genes (~10³ • Affinity:
molecules/cell). o Specific binding (~10²–10³ molecules).
o Affinity Chromatography: ▪ Example: Antibody purification (~10²
molecules).
▪ Specific binding (~10²–10³
• Centrifugation:
molecules).
o Density-based (~10²–10³ samples).
▪ Example: His-tag purification (~10² ▪ Example: Plasmid centrifugation
molecules). (~10² samples).
▪ High specificity (~95–99%). • Efficiency: ~90% separation accuracy.
▪ Example: Specificity ~98%.
5.3 Biological Applications
▪ Molecular Regulation: Affinity genes • Genomics: Purifies ~10⁴ nucleic acids.
(~10³ transcripts/cell). • Proteomics: Isolates ~10³ proteins.
▪ Example: Ligand genes (~10³ • Biotechnology: Enhances ~10² products.
molecules/cell). • Modeling: Optimizes ~10² separation systems.
ToppersNotes / 9614-828-828 84
 Solution: Chromatography.
Answer: A.
Tip: Protein = chromatography.
(2019):
4. What measures RNA purity?
(A) A260/A280 (B) Species count
(C) Both (D) None.
Solution: A260/A280.
Answer: A.
Tip: Purity = A260/A280.
(2020):
5. What is plasmid DNA isolation?
(A) Alkaline lysis (B) Fossils
(C) Both (D) None.
Diagram 4: Separation Methods Solution: Alkaline lysis.
[Description: A diagram showing separation Answer: A.
methods (ion-exchange, affinity, Tip: Plasmid = alkaline.
centrifugation). Mechanisms (SEP genes, (2020):
binding), regulation (H3K27me3), and 6. What characterizes ion-exchange
applications (genomics) are depicted. A side chromatography?
panel illustrates chromatography columns and (A) Charge-based (B) Fossils
centrifugation rotors, with biological roles (e.g., (C) Both (D) None.
biomolecule isolation).] Solution: Charge-based.
Answer: A.
PYQ Analysis Tip: Ion-exchange = charge.
Below are 25 PYQs from CSIR NET Life Sciences (2021):
(2018–2024) related to molecular biology and 7. What regulates RNA extraction?
recombinant DNA methods (Part 1). (A) RNA genes (B) DNA genes
(2018): (C) Both (D) None.
1. What is RNA isolation? Solution: RNA genes.
(A) Extraction (B) Fossils Answer: A.
(C) Both (D) None. Tip: RNA = genes.
Solution: Extraction. (2021):
Answer: A. 8. What shapes protein yield?
Tip: RNA = extraction. (A) Sonication (B) Fossils
(2018): (C) Both (D) None.
2. What drives DNA purification? Solution: Sonication.
(A) Phenol-chloroform (B) Fossils Answer: A.
(C) Both (D) None. Tip: Protein = sonication.
Solution: Phenol-chloroform. (2022):
Answer: A. 9. What drives centrifugation?
Tip: DNA = phenol. (A) Sedimentation (B) Fossils
(2019): (C) Both (D) None.
3. What is protein purification? Solution: Sedimentation.
(A) Chromatography (B) Fossils Answer: A.
(C) Both (D) None. Tip: Centrifugation = sedimentation.
ToppersNotes / 9614-828-828 85
(2022): (2021):
10. What characterizes affinity chromatography? 17. What shapes TRIzol extraction?
(A) Specific binding (B) Fossils (A) Phase separation (B) Fossils
(C) Both (D) None.
(C) Both (D) None.
Solution: Specific binding.
Answer: A. Solution: Phase separation.
Tip: Affinity = binding. Answer: A.
(2023): Tip: TRIzol = separation.
11. What enhances automated extraction? (2020):
(A) Magnetic beads (B) Soil pH 18. What measures purification fold?
(C) Both (D) None.
(A) Fold (B) Species count
Solution: Magnetic beads.
Answer: A. (C) Both (D) None.
Tip: Automated = beads. Solution: Fold.
(2023): Answer: A.
12. What shapes genomic DNA isolation? Tip: Fold = purification.
(A) CTAB (B) Fossils (2019):
(C) Both (D) None.
19. What regulates DNA extraction?
Solution: CTAB.
Answer: A. (A) DNA genes (B) PRO genes
Tip: Genomic = CTAB. (C) Both (D) None.
(2024): Solution: DNA genes.
13. What regulates protein purification? Answer: A.
(A) PRO genes (B) RNA genes Tip: DNA = genes.
(C) Both (D) None.
(2018):
Solution: PRO genes.
Answer: A. 20. What shapes ultracentrifugation?
Tip: PRO = proteins. (A) High speed (B) Fossils
(2024): (C) Both (D) None.
14. What drives RNA precipitation? Solution: High speed.
(A) Ethanol (B) Fossils Answer: A.
(C) Both (D) None.
Tip: Ultra = speed.
Solution: Ethanol.
Answer: A. (2022):
Tip: RNA = ethanol. 21. What drives His-tag purification?
(2023): (A) Affinity (B) Fossils
15. What shapes plasmid purity? (C) Both (D) None.
(A) Column-based (B) Fossils Solution: Affinity.
(C) Both (D) None.
Answer: A.
Solution: Column-based.
Answer: A. Tip: His-tag = affinity.
Tip: Plasmid = column. (2023):
(2022): 22. What enhances RNA-seq preparation?
16. What enhances protein separation? (A) RNA isolation (B) Soil pH
(A) Ion-exchange (B) Soil pH (C) Both (D) None.
(C) Both (D) None.
Solution: RNA isolation.
Solution: Ion-exchange.
Answer: A. Answer: A.
Tip: Protein = ion-exchange. Tip: RNA-seq = isolation.

ToppersNotes / 9614-828-828 86
(2024): 3. Eliminate Incorrect Options:
23. What shapes protein precipitation? o For RNA, match extraction (e.g., TRIzol ≠
(A) Ammonium sulfate (B) Fossils fossils).
(C) Both (D) None. o For separation, match chromatography
Solution: Ammonium sulfate. (e.g., ion-exchange ≠ species count).
Answer: A. 4. Avoid Pitfalls:
Tip: Protein = sulfate. o Don’t confuse genomic (CTAB) vs.
(2021): plasmid (alkaline lysis) DNA.
24. What regulates centrifugation? o Don’t mix up ion-exchange (charge) vs.
(A) SEP genes (B) RNA genes affinity (specific) chromatography.
(C) Both (D) None. o Distinguish TRIzol (RNA) vs. sonication
Solution: SEP genes.
(protein) lysis.
Answer: A.
5. Time Management:
Tip: SEP = centrifugation.
o Allocate 1–2 minutes for Part B
(2020):
questions (e.g., RNA isolation
25. What enhances cloning preparation?
definition).
(A) DNA isolation (B) Fossils
o Spend 3–4 minutes for Part C questions
(C) Both (D) None.
(e.g., purification fold calculation).
Solution: DNA isolation.
o Practice sketching extraction workflows
Answer: A.
Tip: Cloning = DNA. and chromatography setups.

Exam Tips Molecular Biology and Recombinant

1. Memorize Key Facts: DNA Methods - Part 2
o RNA Isolation: TRIzol, column-based
1. Overview of Molecular Biology and
(~10²–10³ samples, e.g., mRNA), purity
Recombinant DNA Methods - Part 2
(~A260/A280 ~2.0).
Gel electrophoresis and molecular cloning
o DNA Isolation: Genomic (CTAB, ~10²
are cornerstone techniques in molecular
μg/g), plasmid (alkaline lysis, ~1.8
biology, enabling the analysis and
A260/A280).
o Protein Purification: Sonication, affinity manipulation of RNA, DNA, and proteins.
chromatography (~10² mg/g, ~10² fold). Gel electrophoresis separates biomolecules
o Separation: Ion-exchange (charge, ~95% based on size, charge, or isoelectric point,
resolution), centrifugation (~10² S, ~90% supporting ~10⁷ proteomic and genomic
purity). studies globally, while molecular cloning
o Regulation: RNA (RNase inhibitors), DNA introduces DNA or RNA fragments into
(polymerase), PRO (protease inhibitors), vectors for replication or expression, driving
SEP (binding). ~10⁶ recombinant DNA applications. Part 2
o Examples: Indian rice RNA (~10⁵ μg), explores 1D and 2D gel electrophoresis,
plasmid DNA (~10⁵ μg). isoelectric focusing (IEF), and cloning in
2. Master Numericals: bacterial (e.g., E. coli) and eukaryotic (e.g.,
o Calculate RNA yield (e.g., ~10² μg/g yeast, mammalian cells) systems, critical for
liver). India’s ~10⁷ biotechnology and research
o Estimate protein purification fold (e.g., sectors.
~10² for insulin). • Gel Electrophoresis:
o Compute centrifugation g-force (e.g., o 1D, 2D, IEF (~10²–10³ techniques).
~10⁵ g for ultracentrifugation).
ToppersNotes / 9614-828-828 87