Experimental Biochemistry, Series-I

  1. 1)      General Lab Safety                                                         7 modules

    2)      Solutions, pH, Buffers & Dilution Series                     33 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                                                         12 experiments

    		 

Purchase Information

 A Practical Manual of Experimental Biochemistry and Biotechnology (Series I)-BLUE


Price: 600/- PKR
Edition: 2026-2027 (4th edition)
ISBN: 978-627-7502-21-8 (Print)
Published by Dr. Muhammad Ali

For Public and Private Universities of Pakistan

Seven (07) 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 will be 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

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Experimental Biochemistry and Biotechnology
Experimental Biochemistry
Sample Pages of the Manual (Experimental Biochemistry and Biotechnology)
Your can send email to ask for excel sheet of calculations of any experiment. [email protected]
This manual is written according to modern education system and objective based learning.

Contents of EBB Series-I (Blue) 4th edition; 2026

 

  1. Safety Rules for Life Sciences and Chemistry Laboratory 1

1.1 Instructions for practical work in the laboratory             1

1.2 Understanding the precautions of laboratory safety   1

1.3 RAMP: A framework for laboratory safety            2

1.4 First aid in the laboratory               2

1.5 First-aid kit in the laboratory        3

1.6 Personal and general laboratory safety 3

1.7 Symbols of laboratory hazards and PPE              4

  1. Solutions, pH, Buffers, and Dilution Series 5

Experiment 2.1- Prepare 50 mL of 0.2 molar (0.2 M) glucose solution.    7

Experiment 2.2- Using 50 g of water and make 0.5 molal NaCl solution.               8

Experiment 2.3- Calculate molarity of concentrated sulfuric acid of 95 % purity (ρ =1.84 g/mL).         8

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.           9

Experiment 2.5- How will you make 500 mL of 0.601 M solution of HNO3 (density 1.48 g/mL)?             10

Experiment 2.6- How will you prepare 500 mL of 1 M H3PO4 solution when it is 85.0 % pure. 10

Experiment 2.7- Prepare 150 mL of 2 % (w/v) NaCl solution.         11

Experiment 2.8- What is w/v % of 1 M NaCl solution?        12

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?     12

Experiment 2.10- How to make 100 µL of 2 mM dNTPs (dATP, dTTP, dCTP, and dGTP) from 100 mM stock of each?            13

Experiment 2.11- Two lyophilized PCR primers are received and it is written 50 and 49 nano-moles. How will you make 100 µmolar stock and 10 µmolar working solution?            13

Experiment 2.12- How will you design a PCR reaction and what is the final concentration of primer, buffer, MgCl2, dNTPs, and polymerase?      14

Experiment 2.13- Prepare 80 mL of 70 % (v/v) ethanol (C2H5 -OH) solution.      15

Experiment 2.14- Prepare 200 mL of 200-ppm CaCl2 solution.   15

Experiment 2.15- Prepare 100 mL of 20 ppb CaCl2 solution from its 100 ppm stock solution.               16

Experiment 2.16- A certain solution has 1 × 10-5 M CaCl2. What is the concentration in ppm?             16

Experiment 2.17- How much absolute ethanol will be added to 50 mL of 70 % ethanol to make its 80 %?                17

Experiment 2.18- 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 2.19- Prepare serial and parallel dilutions of 1 M CuSO4 solution. 18

Experiment 2.20- Prepare 200 mL of 0.5 M Tris buffer and adjust pH to 6.8 (pKa of Tris = 8.1). 19

Experiment 2.21- Prepare 200 mL of 0.5 M EDTA solution and adjust pH to 8.00.            20

Experiment 2.22- Prepare 50 mL of 10 % SDS solution.    20

Experiment 2.23- 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. 21

Experiment 2.24- Prepare 1 liter of 0.2 M phosphate buffer of 7.3 pH.     22

Experiment 2.25- Prepare acetate and phosphate buffers of different pH.           23

Experiment 2.26- Prepare 100 mL of lysis buffer for protein extraction.  23

Experiment 2.27- Prepare 40 mL of DNA extraction buffer.             24

Experiment 2.28- Demonstration of diffusion and dialysis              25

Experiment 2.29- Electrolytes and non-electrolytes            26

Experiment 2.30- How will you measure pH by pH meter?              27

Experiment 2.31- Donnan equilibrium          28

Experiment 2.32- Compare calculated and measured pH values of a solution that has 10-7 moles of HCl dissolved per mL of solution.    30

Experiment 2.33- 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

  1. 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 reducing sugars (aldo and keto)         42

Experiment 3.9- Seliwanoff’s test for the identification of keto group      43

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- Quantitation 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- Demonstration of Glucose Tolerance Test (GTT)            51

  1. Biochemistry of Amino Acids and Proteins 52

Experiment 4.1- Perform different qualitative tests for the identification of amino acids            53

Experiment 4.2- Draw titration curve of glycine and mark pI and pKa 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 (Bicinchoninic acid) 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 molar mass of proteins by SDS-PAGE            68

Experiment 4.18- Electrophoretic Mobility Shift Assay (EMSA)     70

  1. Biochemistry of Enzymes 71

Experiment 5.1- Prepare LB media and LB agar plates for bacterial culture.        73

Experiment 5.2- Isolate pure colonies of E. coli (DH10-α) from its culture.           74

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 β-galactosidase (lactase) activity          88

Experiment 5.14- Find optimum temperature for enzymatic activity of trypsin. 91

Experiment 5.15- Determine optimum pH for trypsin activity (colorimetric).      92

Experiment 5.16- Determine the optimum pH of chymotrypsin colorimetrically.             93

Experiment 5.17- Determine substrate specificity of trypsin and chymotrypsin.             95

Experiment 5.18- Determine the optimum pH for the activity of pepsin.               96

Experiment 5.19- Find Km value of pepsin enzyme.             97

Experiment 5.20- Effect of temperature on the activity of catalase enzyme         99

Experiment 5.21- Determination of thermal stability, deactivation kinetics, and half-life of α-amylase                99

Experiment 5.22- Immobilization of enzymes using different supports and techniques              104

Experiment 5.23- Characterization of free and immobilized lactase enzymes   105

  1. 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 value 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            113

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- Liebermann-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

  1. Proximate Analysis 120

Experiment 7.1- Determination of moisture content of raisin        120

Experiment 7.2- Determination of ash content of cabbage             121

Experiment 7.3- Crude fiber determination of milled rice 121

Experiment 7.4- Extraction and quantitation of soluble sugars (phenol-sulfuric acid)  122

Experiment 7.5- Extraction and quantitation of starch from plant tissue               124

Experiment 7.6- Extraction of oil by Soxhlet method (Ether-mediated extraction of lipids)        126

Experiment 7.7- Crude and true protein quantification of rice sample by Kjeldahl’s method   127

Experiment 7.8- Estimation of fat content in almond          128

Experiment 7.9- Estimation of fat by chloroform–methanol procedure (Folch extraction) and % of ether free extract      129

Experiment 7.10- Estimation of vitamin C (Ascorbic acid) in orange samples (mg /g of fruit)    130

Experiment 7.11- Estimation of vitamin A with trichloroacetic acid           131

Experiment 7.12- Estimation of casein and lactose in milk             132

  1. Appendices 134

8.1- Reagents for Biochemistry and Biotechnology lab      134

8.2- Composition for Tris-glycine SDS-Polyacrylamide Gel Electrophoresis (SDS-PAGE)           135

  1. Glossary 136
  2. Bibliography and further reading 139
  3. Rubrics and Instructors’ Guide 142
  4. Relevancy to industry /Job market 144
  5. Benchmarking with national and international industry 144
MMGL- Life sciences
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Fulfilling contents of courses from world top rank universities

  1. 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

  1. Harvard University

o MCB-101 — Molecular and Cellular Biology Laboratory

  1. Yale University

o     MB&B 251L — Laboratory for Biochemistry and Biophysics

  1. Johns Hopkins University

o     AS.020.315 — Biochemistry Laboratory

  1. Cornell University

o     BIOMG 4400 — Laboratory in Biochemistry and Molecular Biology

  1. UC Berkeley

o     MCB 110L — General Biochemistry and Molecular Biology Laboratory

  1. UCLA

o     CHEM 153L — Biochemical Methods Laboratory

  1. University College London (UCL), UK

o     BIOC0006 — Practical Biochemistry

  1. Imperial College London, UK

o     LIFE40003 — Biological Chemistry and Laboratory Techniques

  1. University of Edinburgh, UK

o    BILG08014 — Biochemistry Laboratory Methods

  1. University of Toronto, Canada

o     BCH370H1 — Laboratory Course in Biochemistry

  1. University of British Columbia (UBC), Canada

o     BIOC 301 — Biochemistry Laboratory

  1. McGill University, Canada

o     BIOC 312 — Biochemistry Laboratory

  1. University of Wisconsin-Madison

o     BIOCHEM 551 — Biochemical Methods

  1. University of Texas at Austin

o     BCH 369L — Biochemistry Laboratory

  1. Purdue University

o     BCHM 30200 — Analytical Biochemistry Laboratory

  1. The University of Melbourne (UniMelb)

o     BIOM10002– Biomedical Science Research Core

o     BCMB20005– Techniques in Molecular Science.

o     BCMB30010– Advanced Practical Molecular Science

  1. University of Sydney (USYD)

o     BIOL1007– From Molecules to Ecosystems

o     BCMB2001 / BCMB2901- Advanced Biochemistry and Molecular Biology

o     AGCH3026- Food Chemistry and Biochemistry

  1. 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).

  1. The University of Queensland (UQ)

o     BIOL1020- Genes, Cells & Evolution

o     BIOC2000- Biochemistry & Molecular Biology

o     FOOD2000– Food Science

  1. University of New South Wales (UNSW)

o     BIOC2101– Principles of Biochemistry

  1. KAIST, South Korea

o     BS202: Biochemistry I & BS301: Biochemistry II

  1. 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

  1. University of British Columbia (UBC)

o     BIOC301- Biochemistry Laboratory

o     BIOC421 or BIOC460- Recombinant DNA Techniques / Advanced Techniques in Biochemistry

  1. University of Guelph

o     BIOC*3570- Analytical Biochemistry

o     FOOD*2150 or FOOD*3030- Introduction to Food and Nutritional Sciences / Food Chemistry 1

  1. University of Ottawa (uOttawa)

o     BCH 3346- Biochemistry Laboratory II

o     BCH 3356- Molecular Biology Laboratory

  1. King Saud University

o     BCH 201- General Biochemistry-I

  1. Qatar University

o     BIOM 201- Course Title: Medical Biochemistry

o     NUTR 321- Food Chemistry

  1. United Arab Emirates University (UAEU)

o     BIOC 205- Principles of Biochemistry

  1. The American University in Cairo (AUC)

o     CHEM 311- Biochemistry I Lab

  1. Sultan Qaboos University

o     BIOL 3010- General Biochemistry

 

This manual fits well for BS Biochemistry, BS Chemistry, BS Biotechnology and Allied Life sciences at

  1. Quaid-i-Azam University (QAU)
  2. International Islamic University, Islamabad (IIUI)
  3. National University of Sciences and Technology (NUST)
  4. COMSATS University Islamabad (CUI)
  5. University of the Punjab (PU), Lahore
  6. University of Agriculture, Faisalabad (UAF)
  7. Government College University (GCU), Lahore
  8. Government College University (GCUF), Faisalabad
  9. The University of Faisalabad
  10. Bahauddin Zakariya University (BZU), Multan
  11. University of Sargodha (UOS)
  12. The Islamia University of Bahawalpur (IUB)
  13. University of Veterinary and Animal Sciences (UVAS), Lahore
  14. University of Gujrat (UOG)
  15. University of Education (UE), Lahore
  16. Lahore College for Women University (LCWU), Lahore
  17. Fatima Jinnah Women University (FJWU), Rawalpindi
  18. University of Jhang (UOJ)
  19. Lahore University of Management Sciences (LUMS), Lahore
  20. Forman Christian College (FCCU), Lahore
  21. University of Central Punjab (UCP), Lahore
  22. University of Management and Technology (UMT), Lahore
  23. The University of Lahore (UOL)
  24. Superior University, Lahore
  25. Riphah International University (Faisalabad and Lahore Campuses)
  26. GIFT University
  27. The University of Chenab, Gujrat
  28. Superior University
  29. Ali Institute of Education
  30. Qarshi University
  31. University of Management and Technology
  32. National College of Business Administration and Economics
  33. Global Institute
  34. Imperial College
  35. University of Lahore
  36. Lahore Leads University
  37. Beaconhouse National University
  38. Hajvery University
  39. University of South Asia
  40. Lahore Institute of Science and Technology, Lahore.
  41. Lahore Garrison University
  42. NUR International University
  43. Green International University
  44. Rashid Latif Khan University
  45. Institute of Southern Punjab
  46. Times Institute, Multan
  47. Multan University of Science and Technology
  48. University of Wah
  49. HITEC University
  50. South Punjab Institute of Science and Technology D.G. Khan
  51. Ghazi National Institute of Engineering and Sciences
  52. University of Sialkot (USKT)
  53. Minhaj University, Lahore
  54. University of Karachi (UoK) — Karachi
  55. University of Sindh (UoS) — Jamshoro
  56. Shah Abdul Latif University (SALU) — Khairpur
  57. Federal Urdu University of Arts, Sciences & Technology (FUUAST) — Karachi Campus
  58. Dow University of Health Sciences (DUHS) — Karachi
  59. People’s University of Medical & Health Sciences for Women (PUMHSW) — Shaheed Benazirabad (Nawabshah)
  60. Muhammad Ali Jinnah University (MAJU) — Karachi
  61. Jinnah University for Women (JUW) — Karachi
  62. Ziauddin University — Karachi
  63. Sohail University — Karachi
  64. Dadabhoy Institute of Higher Education (DIHE) — Karachi
  65. Ilma University (formerly Biztek) — Karachi
  66. University of Balochistan (UoB), Quetta
  67. University of Turbat (UoT), Turbat
  68. Sardar Bahadur Khan (SBK) Women’s University, Quetta
  69. BUITEMS, Quetta
  70. Al-Hamd Islamic University, Quetta
  71. University of Peshawar (UoP), Peshawar
  72. The University of Agriculture, Peshawar
  73. Abdul Wali Khan University Mardan (AWKUM)
  74. Hazara University, Mansehra
  75. The University of Haripur
  76. Gomal University, Dera Ismail Khan
  77. Shaheed Benazir Bhutto Women University (SBBWUP), Peshawar
  78. Kohat University of Science and Technology (KUST), Kohat
  79. University of Swat, Swat
  80. Khushal Khan Khattak University, Karak
  81. Women University Swabi, Swabi
  82. University of Malakand (UOM), Chakdara
  83. CECOS University of IT and Emerging Sciences, Peshawar
  84. Qurtuba University of Science & Information Technology, Peshawar & D.I. Khan Campuses
  85. Abasyn University, Peshawar
  86. Sarhad University of Science and Information Technology (SUIT), Peshawar
  87. City University of Science and Information Technology (CUSIT), Peshawar
  88. Northern University, Nowshera
  89. Ghazali Institute of Medical Sciences (GIMS), Peshawar
  90. ​
  91.  
Contents of Series-I
Detail of Series-I
Contents of Series-II
Detail of Series-II
Contents of Series-III
Detail of Series-III