1) General Lab Safety 5 modules
2) Solutions and Buffers 34 experiments
3) Biochemistry of Carbohydrates 18 experiments
4) Biochemistry of Amino Acids & Proteins 18 experiments
5) Biochemistry of Enzymes 23 experiments
6) Biochemistry of Lipids 15 experiments
7) Proximate Analysis 11 experiments
Purchase Information
A Practical Manual of Experimental Biochemistry and Biotechnology (Series I)-BLUE
Price: 700/- PKR
Edition: 2026-2027 (4th edition)
ISBN: 978-627-7502-21-1 (Print)
Published by Dr. Muhammad Ali
For all Public and Private Universities of Pakistan
Five (05) modules of lab safety and One hundred and eleven (119) full-length experiments are included.
© Copyright 2026. All rights reserved by Dr. Muhammad Ali
Contact us for purchase and online ordering:
0300-6602034
[email protected]
This edition is available on Amazon, Draz, and several other international stores.
Video lectures of almost all the experiments will be uploaded sequentially.
All rights of this experimental manual is with Dr. Muhammad Ali
Contents of EBB Series-I (Blue) edition of 2026
Safety Rules for Lifesciences and Chemistry Laboratory. 1
1.1 Instructions for practical work in the laboratory. 1
1.2 Understanding the precautions of lab safety. 1
1.3 First aid in the laboratory. 1
1.4 First aid kit in the laboratory. 2
1.5 Personal and general lab safety. 2
Solutions and Buffers. 4
Experiment 2.1- Prepare 50 mL of 0.2 molar (0.2 m) glucose solution. 6
Experiment 2.2- Using 50 g of water and make 0.5 molal NaCl solution. 7
Experiment 2.3- Calculate molarity of concentrated sulfuric acid of 95% purity (ρ =1.84 g/mL). 7
Experiment 2.4- Prepare 100 mL of 0.1 normal (0.1 N) sulfuric acid (H2SO4) solution. Concentrated sulfuric acid has purity and density of 95% and 1.84 g/mL. 8
Experiment 2.5- How will you make 500 mL of 0.601 m solution of HNO3 (density 1.48 g/mL)?. 9
Experiment 2.6- How will you prepare 500 mL of 1 m H3PO4 solution when it is 85.0% pure. 9
Experiment 2.7- Prepare 150 mL of 2% (w/v) NaCl solution. 10
Experiment 2.8- What is w/v % of 1 m NaCl solution?. 11
Experiment 2.9- How much of 100 mm dNTPs will be added to 100 µL of 2 mm dNTPs solution to make its 2.5 mm?. 11
Experiment 2.10- How will you make 100 µL of 2 mm dNTPs (dATP, dTTP, dCTP, and dGTP) from 100 mm stock of each?. 12
Experiment 2.11- Two lyophilized PCR primers are received and it is written 50 and 48 nano-moles. How will you make 100 µmolar stock and 10 µmolar working solution?. 12
Experiment 2.12- How will you design a PCR reaction and what is the final concentration of primer, buffer, MgCl2, dNTPs, and polymerase?. 13
Experiment 2.13- A spherical bacterium has a diameter of 1 µm and contains three molecules of a particular protein. What is molar concentration of the protein?. 14
Experiment 2.14- Prepare 80 mL of 70% (v/v) ethanol (C2H5 -OH) solution. 14
Experiment 2.15- Prepare 200 mL of 200-ppm CaCl2 solution. 15
Experiment 2.16- Prepare 100 mL of 20 ppb CaCl2 solution from its 100 ppm stock solution. 15
Experiment 2.17- A certain solution has 1 × 10-5 m CaCl2. What is the concentration in ppm?. 16
Experiment 2.18- How much absolute ethanol will be added to 50 mL of 70% ethanol to make its 80%?. 17
Experiment 2.19- What will be final concentration of NaCl solution if 6 mL of 5% NaCl solution is mixed with 7 mL of 3.2% NaCl solution?. 17
Experiment 2.20- Prepare serial and parallel dilutions of 1 m CuSO4 solution. 18
Experiment 2.21- Prepare 200 mL of 0.5 m Tris buffer and adjust pH to 6.8 (pK of Tris = 8.1). 19
Experiment 2.22- Prepare 200 mL of 0.5 m EDTA solution and adjust pH to 8.00. 19
Experiment 2.23- Prepare 50 mL of 10 % SDS solution. 20
Experiment 2.24- Ten grams (10 g) of each of K2HPO4, and KH2PO4 were dissolved in 500 mL of water; show difference between calculated and measured pH values. 20
Experiment 2.25- Prepare 1 liter of 0.2 m phosphate buffer of 7.3 pH. 21
Experiment 2.26- Prepare acetate and phosphate buffers of different pH. 23
Experiment 2.27- Prepare 100 mL of lysis buffer for protein extraction. 23
Experiment 2.28- Prepare 40 mL of DNA extraction buffer. 24
Experiment 2.29- Demonstration of diffusion and dialysis. 25
Experiment 2.30- Electrolytes and non-electrolytes. 26
Experiment 2.31- How will you measure pH by pH meter?. 27
Experiment 2.32- Donnan equilibrium and osmosis. 28
Experiment 2.33- Compare calculated and measured pH values of a solution that has 10-7 moles of HCl dissolved per mL of solution. 30
Experiment 2.34- What is the final pH of the solution if 1 mL of 0.2 molar solution of NaOH is added to make 500 mL of solution?. 31
Exam questions for solutions, buffers, and pH.. 31
Biochemistry of Carbohydrates. 34
Experiment 3.1- Molisch’s test for the identification of carbohydrates. 37
Experiment 3.2- Iodine test for the identification of polysaccharide. 37
Experiment 3.3- Fehling’s test for the identification of reducing sugars. 38
Experiment 3.4- Benedict’s test for the identification of reducing sugar. 39
Experiment 3.5- Estimation of optical activity of D-glucose by polarimetry. 39
Experiment 3.6- Quantitation of sucrose by polarimeter. 40
Experiment 3.7- Barfoed’s test for the identification of monosaccharides. 41
Experiment 3.8- Osazone’s test for aldo and keto group of sugar. 42
Experiment 3.9- Seliwanoff’s test for the identification of keto group. 42
Experiment 3.10- Bial’s test for the identification of pentoses. 43
Experiment 3.11 Quantitation of glucose by using DNS reagent. 44
Experiment 3.12- Estimation of blood glucose by glucose oxidase method. 45
Experiment 3.13- Chromatographic separation of monosaccharides. 46
Experiment 3.14- HCl-mediated hydrolysis of sucrose. 48
Experiment 3.15- HCl-mediated hydrolysis of starch. 48
Experiment 3.16- Rapid extraction of glycogen from fresh mouse liver. 49
Experiment 3.17- HCl-mediated hydrolysis of glycogen obtained from mouse liver. 50
Experiment 3.18- Glucose tolerance test (GTT) for the diagnosis of diabetes. 51
Biochemistry of Amino Acids and Proteins. 52
Experiment 4.1- Perform different qualitative test for the identification of amino acids. 53
Experiment 4.2- Draw titration curve of glycine and mark pI and values. 54
Experiment 4.3- Hydrolysis of protein from dried plant tissues. 55
Experiment 4.4- Qualitative test for amino acids obtained by the hydrolysis of proteins. 55
Experiment 4.5- Extraction of total proteins from leaves of rice plant. 57
Experiment 4.6- How will you extract total proteins from animal tissues?. 57
Experiment 4.7- Separation of a nuclear and cytosolic fraction of proteins. 58
Experiment 4.8- Circular chromatography. 59
Experiment 4.9- Fractionation of color pigments of berseem.. 60
Experiment 4.10- Absorption spectra (wavelength scan) of albumin to find its wavelength of maximum absorption. 61
Experiment 4.11- Protein quantitation by UV spectroscopy. 62
Experiment 4.12- Estimation of total serum proteins by the Biuret method. 63
Experiment 4.13- Estimation of total serum proteins by Lowry assay. 64
Experiment 4.14- Quantitation of proteins by BCA assay. 65
Experiment 4.15- Bradford assay (dye binding assay) for protein quantitation. 66
Experiment 4.16- Size exclusion (column) chromatography for the separation of proteins. 67
Experiment 4.17- Determination of molecular weight of proteins by SDS-PAGE. 68
Experiment 4.18- Electrophoretic Mobility Shift Assay (EMSA). 70
Biochemistry of Enzymes. 71
Experiment 5.1- Prepare nutrient LB-media and LB-agar plates for bacterial culture. 73
Experiment 5.2- Isolate pure colonies of E. coli (DH10-α) from its culture. 73
Experiment 5.3- Transformation of plasmid vectors in competent cells (E. coli). 74
Experiment 5.4- Purification of GST-tagged enzyme from bacterial culture. 75
Experiment 5.5- Microbial technology for the production of lactase (β-Galactosidase). 77
Experiment 5.6- Purify enzymes by ammonium sulfate precipitation, dialysis, and gel filtration chromatography 79
Experiment 5.7- Purification of enzymes by ion exchange chromatography. 82
Experiment 5.8- Produce cellulase enzyme using Bacillus subtilis (AU-1). 84
Experiment 5.9- Hydrolysis of starch using amylase enzyme. 85
Experiment 5.10- Effect of pH on the enzymatic activity of salivary amylase. 85
Experiment 5.11- Effect of temperature on the reaction rate of α-amylase. 86
Experiment 5.12- Effect of cofactors and metal ions on enzyme (lactase) activity. 87
Experiment 5.13- Effect of inhibitors on beta-galactosidase (lactase) activity. 89
Experiment 5.14- Find optimum temperature for enzymatic activity of trypsin. 91
Experiment 5.15- Determine optimum pH for trypsin activity (calorimetric). 92
Experiment 5.16- Determine the optimum pH of chymotrypsin calorimetrically. 94
Experiment 5.17- Determine substrate specificity of trypsin and chymotrypsin. 96
Experiment 5.18- Determine the optimum pH for the activity of pepsin. 97
Experiment 5.19- Find Km value of pepsin enzyme. 98
Experiment 5.20- Effect of temperature on the activity of catalase enzyme. 99
Experiment 5.21- Find thermodynamic parameters and half-life of amylase. 100
Experiment 5.22- Immobilization of enzymes using different supports and techniques. 103
Experiment 5.23- Characterization of free & immobilized lactase enzymes. 104
Biochemistry of Lipids. 107
Experiment 6.1- Solubility test of lipids in water and organic solvent. 108
Experiment 6.2- Grease spot test for lipids. 108
Experiment 6.3- Test for the identification of free fatty acids in corn oil 109
Experiment 6.4- Determination of iodine number of cooking oil 109
Experiment 6.5- Emulsification of lipids by using bile salt. 110
Experiment 6.6- Fractionation of lipids by using 2-dimensional TLC. 111
Experiment 6.7- Tests for the unsaturation of fatty acids. 112
Experiment 6.8- Saponification value of corn oil (lipid). 113
Experiment 6.9- Extraction of total lipids from mouse brain tissue. 114
Experiment 6.10- Isolation of free fatty acids from soap. 114
Experiment 6.11- Acrolein test for glycerol identification. 115
Experiment 6.12- Dichromate test for the identification of glycerol 115
Experiment 6.13- Liberman-Burchard test for the quantitation of cholesterol 116
Experiment 6.14- Salkowski’s test for cholesterol identification. 117
Experiment 6.15- Colorimetric determination of phospholipids (ammonium ferrothiocyanate method). 118
Proximate Analysis. 120
Experiment 7.1- Determination of moisture contents of raisin. 120
Experiment 7.2- Determination of ash content of cabbage. 121
Experiment 7.3- Extraction and quantitation of soluble sugars (phenol-sulfuric acid). 121
Experiment 7.4- Extraction and quantitation of starch from plant tissue. 124
Experiment 7.5- Extraction of oil by Soxhlet method. 125
Experiment 7.6- Protein quantification by Kjeldahl’s method. 126
Experiment 7.7- Estimation of fat contents in almond. 127
Experiment 7.8- Estimation of fat by chloroform–methanol procedure (Folch extraction). 128
Experiment 7.9- Estimation of vitamin C (Ascorbic acid) in orange samples (mg /g of fruit). 129
Experiment 7.10- Colorimetric estimation of vitamin A with trichloroacetic acid. 130
Experiment 7.11- Estimation of casein and lactose in milk. 131
Appendices. 133
8.1- Reagents for Biochemistry and Biotechnology lab. 133
8.2- Composition for Tris-glycine SDS-Polyacrylamide Gel Electrophoresis (SDS-PAGE). 134
Bibliography and further reading. 135
Experiment 3.1- Molisch’s test for the identification of carbohydrates
1- Sucrose shows red color (tube A). 2- The appearance of deep blue color will indicate the presence of polysaccharides (tube B). 3- Glucose shows yellowish color (tube C).
Experiment 3.2- Iodine test for the identification of polysaccharide
Deep blue coloration that disappear on heating and appears on cooling shows polysaccharises
Experiment 2.13- Prepare serial and parallel dilutions of 1 M CuSO4 solution.
Serial dilutions are used to find IC_50 values of drugs. Parallel dilutions can be used to make a standard for the calibration curve. The dilution series can also be used for spiking.
Experiment 4.2- Draw titration curve of glycine and mark pI and 〖pK〗_(a )values.
Glycine has pKa1, pKa2, and pI values of 2.34, 9.6, and 5.97 respectively.
Experiment 5.3- Transformation of plasmid vectors in competent cells (E. coli)
Transformed bacteria are used in enzymology for the production of enzymes. Moreover, the transformation is used to multiply copies of plasmids containing genes of our interest.
Fulfilling contents of courses from world top rank universities
- MIT (Massachusetts Institute of Technology)
o 7.002— Fundamentals of Experimental Biology (Core experimental biochemistry, handling assays, and cell measurements)
o 7.02— Experimental Biology and Communication
- Harvard University
o MCB-101 — Molecular and Cellular Biology Laboratory
- Yale University
o MB&B 251L — Laboratory for Biochemistry and Biophysics
- Johns Hopkins University
o AS.020.315 — Biochemistry Laboratory
- Cornell University
o BIOMG 4400 — Laboratory in Biochemistry and Molecular Biology
- UC Berkeley
o MCB 110L — General Biochemistry and Molecular Biology Laboratory
- UCLA
o CHEM 153L — Biochemical Methods Laboratory
- University College London (UCL), UK
o BIOC0006 — Practical Biochemistry
- Imperial College London, UK
o LIFE40003 — Biological Chemistry and Laboratory Techniques
- University of Edinburgh, UK
o BILG08014 — Biochemistry Laboratory Methods
- University of Toronto, Canada
o BCH370H1 — Laboratory Course in Biochemistry
- University of British Columbia (UBC), Canada
o BIOC 301 — Biochemistry Laboratory
- McGill University, Canada
o BIOC 312 — Biochemistry Laboratory
- University of Wisconsin-Madison
o BIOCHEM 551 — Biochemical Methods
- University of Texas at Austin
o BCH 369L — Biochemistry Laboratory
- Purdue University
o BCHM 30200 — Analytical Biochemistry Laboratory
- The University of Melbourne (UniMelb)
o BIOM10002– Biomedical Science Research Core
o BCMB20005– Techniques in Molecular Science.
o BCMB30010– Advanced Practical Molecular Science
- University of Sydney (USYD)
o BIOL1007– From Molecules to Ecosystems
o BCMB2001 / BCMB2901- Advanced Biochemistry and Molecular Biology
o AGCH3026- Food Chemistry and Biochemistry
- Monash University
o CHM1011 / CHM1022- Chemistry 1 / Chemistry 2 (Covers the fundamental solution preparation and pH/buffer modules).
o BCH2011- Structure and Function of Cellular Biomolecules
o Course Code: BCH2022– Metabolic Regulation and Cell Signalling or BTE2011 (Introduction to Biotechnology).
- The University of Queensland (UQ)
o BIOL1020- Genes, Cells & Evolution
o BIOC2000- Biochemistry & Molecular Biology
o FOOD2000– Food Science
- University of New South Wales (UNSW)
o BIOC2101– Principles of Biochemistry
- KAIST, South Korea
o BS202: Biochemistry I & BS301: Biochemistry II
- University of Toronto (U of T)
o CHM135H1 / CHM136H1- Chemistry: Physical Principles / Organic Chemistry I
o BCH370H1 (St. George Campus) or BIOC23H3 (Scarborough Campus)- Laboratory Course in Biochemistry / Practical Approaches to Biochemistry
- University of British Columbia (UBC)
o BIOC301- Biochemistry Laboratory
o BIOC421 or BIOC460- Recombinant DNA Techniques / Advanced Techniques in Biochemistry
- University of Guelph
o BIOC*3570- Analytical Biochemistry
o FOOD*2150 or FOOD*3030- Introduction to Food and Nutritional Sciences / Food Chemistry 1
- University of Ottawa (uOttawa)
o BCH 3346- Biochemistry Laboratory II
o BCH 3356- Molecular Biology Laboratory
- King Saud University
o BCH 201- General Biochemistry-I
- Qatar University
o BIOM 201- Course Title: Medical Biochemistry
o NUTR 321- Food Chemistry
- United Arab Emirates University (UAEU)
o BIOC 205- Principles of Biochemistry
- The American University in Cairo (AUC)
o CHEM 311- Biochemistry I Lab
- Sultan Qaboos University
o BIOL 3010- General Biochemistry
This manual fits well for BS Biochemistry, BS Chemistry, BS Biotechnology and Allied Life sciences at
- Quaid-i-Azam University (QAU)
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- National University of Sciences and Technology (NUST)
- COMSATS University Islamabad (CUI)
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- Muhammad Ali Jinnah University (MAJU) — Karachi
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- Ziauddin University — Karachi
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- Ilma University (formerly Biztek) — Karachi
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- The University of Haripur
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- Shaheed Benazir Bhutto Women University (SBBWUP), Peshawar
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- University of Swat, Swat
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- Women University Swabi, Swabi
- University of Malakand (UOM), Chakdara
- CECOS University of IT and Emerging Sciences, Peshawar
- Qurtuba University of Science & Information Technology, Peshawar & D.I. Khan Campuses
- Abasyn University, Peshawar
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- Northern University, Nowshera
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-