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Analytical Techniques
in Biotechnology
(A COMPLETE LABORATORY MANUAL)
 Author's Profile
Goutam Bhowmik received his BSc in Agriculture (1963) and MSc in Agriculture (1965)
from Banaras Hindu University (BHU), Varanasi. He then obtained his PhD in Genetics
and Plant Breeding from BHU, where his field of research was in micro-mutational and
cytogenetical studies on barley.
His first work assignment was as a Lecturer at BHU from 1966 to 1969. From 1970 to
2004, he was a faculty member at Gauhati University, Guwahati (Assam). He has held the
position of lecturer, reader, professor and HOD in the Department of Biotechnology, and
was also a member of the Academic and Executive Council of Gauhati University. Fifteen
students have received doctorate degrees under his supervision and guidance. His research
papers are focused on tissue culture and isolation of mutations in pineapple, orchids, tea, pigeon pea and French
beans. Moreover, he has standardised in-vitro propagation of many medicinal plants.
An eminent academician, Dr Bhowmik has published 45 and 32 research papers in national and international
journals, respectively. He has visited many European countries as a participant of the Cultural Exchange Programme,
Government of India and has conducted many training programmes as well as research projects sponsored by DBT,
ICAR, CSIR, G B Pant Institute and ASTEC (Assam). He was a member and office bearer of various national
scientific societies. He was the organiser of a DBT sponsored national seminar on ‘Patenting in Biotechnology’ and
has also conducted several other seminars in the field of Biotechnology.
His authorial activities include being the adaptation author of a book on Genetics (4th Ed.) and editing a book
on Patenting.

Sujoy Bose is a Lecturer in the Department of Biotechnology, Gauhati University, Guwahati,

Assam; and teaches Cell and Molecular Biology, Genetic Engineering, Microbiology,
Immunology and Medical Biotechnology. He received his MSc degree in Biotechnology in
the year 2003, and after qualifying the CSIR-UGC NET JRF and GATE exams, completed
his PhD in Biosciences. He is a lifetime member of Indian Association of Cancer Research
(IACR), Asia-Pacific Association in Studies on Liver (APASL), European Association on
Studies on Liver (EASL), and North-east Biotech Association (NEBA).
Dr Bose is the author of a number of scientific research papers in peer-reviewed journals
of high impact factor on cancer signaling, Hepatitis B, and antigen presentation machinery.
He has also presented papers in national and international scientific conferences, and has been a recipient of young
investigator awards. He has also worked in institutes of national and international repute like CDFD, Hyderabad,
G B Pant Hospital, New Delhi, and University of Tokyo Hospital, Tokyo University, Japan. He specialises in the
area of molecular biology, cancer biology, virology and is actively involved in educational and research work in
the field of Biomedical Sciences.
Analytical Techniques
in Biotechnology
(A COMPLETE LABORATORY MANUAL)

Goutam Bhowmik
Former Professor and HOD
Department of Biotechnology
Gauhati University, Guwahati

Sujoy Bose
Lecturer
Department of Biotechnology
Gauhati University, Guwahati

Tata McGraw Hill Education Private Limited

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 Contents

Preface xi
Abbreviations xiii

1: Introduction to the Laboratory 1

∑ Basic Pre-Laboratory Preparations 2
∑ Biology Lab Safety Rules 2
∑ Philosophical and Ethical Issues 5
∑ Suggestions to the Instructor 6
∑ General Information for Keeping a Laboratory Notebook 7
∑ Laboratory Reports 7
(a) Aim or Objective 7
(b) Basic Principle in Brief 7
(c) Materials or Requirements 7
(d) Methods or Procedure 7
(e) Data Tabulation or Observation 7
(f) Result(s) or Finding(s) 7
(g) Figure(s) and Finding(s) 7
(h) Discussion and Conclusion(s) 7
(i) Special Precautions (if any) 8
(j) References 8
∑ Ideal Laboratory Conditions 8
∑ Operation Techniques of Some Equipment and their Basic Working Principles 9
(a) Pipetting with Pipetman 9
(b) Measuring Absorbance with a Spectrophotometer 10
(c) Preparing Dilutions 11
(d) Buffer and pH Determination 12
(e) Metric Length and Fluid Volume 12
(f) Using a Microscope 13
(g) Binomial Nomenclature 15
(h) Centrifugation Technique 15
(i) Polymerase Chain Reaction (PCR) 17
viii Contents

(j) Electrophoresis Technique 20

(k) Sterilization Techniques 25
(l) Autoclaving 27
(m) Distillation 28

2: Genetics 31
Expt. 1 Study of various stages of Meiosis and Mitosis calculating Mitotic Index 31
Expt. 2 Preparing Karyotype of human chromosomes from photograph 36
Expt. 3 Problems in Mendel’s Law of Dominance, Segregation and Independent Assortment 38
Expt. 4 C-banding technique of Zea mayz pachytene chromosomes 43
Expt. 5 Problems in Incomplete Dominance Modified Ratios and Multiple Alleles 45
Expt. 6 Problems in Linkage 52
Expt. 7 Problems in Sex-determination 58
Expt. 8 Problems in Quantitative Inheritance 63
Expt. 9 Estimation of Genetic parameters and partitioning of variance 66
Expt. 10 Problems in Probability, Test of Significance (Chi-Test) and Hardy Winberg’s Law 71

3: Microbiology 78
Expt. 1 A. Isolation of bacteria from Curd, Root nodules, Soil and Water samples 78
B. Isolation of bacterial and fungal spores from ‘Air samples’ 81
Expt. 2 Aseptic technique and transfer of microorganisms for pure culture 83
Expt. 3 A. Staining Gram Positive (+ve) and Gram Negative (–ve) bacteria 85
B. Staining bacterial endospore 88
Expt. 4 Determination of bacterial growth curves by
(a) Manual counting (by microscope), and
(b) UV-Spectrophotometer method 89
Expt. 5 Identification of bacteria through Biochemical test and Pigmentations 91
Expt. 6 Uses of Physical and antimicrobial agents to control microorganisms 95
Expt. 7 Determine which type of garbage is Bio-degradable, and which are non-degradable 102
Expt. 8 Isolation and Identification of Non-Pathogenic Fungi 105
Expt. 9 Determine the cell viability by dye-exclusion method 107
Expt. 10 Plaque count by infecting the bacterium Escherichia coli B with its specific
bacteriophage Coliophage T4. (See Chapter VII; Expt. 8) 109

4: Plant Tissue Culture 114

Expt. 1 Preparation of Stock solution for MS basal medium and Plant Growth Regulator Stock 114
Expt. 2 Aseptic culture technique for establishment and maintenance of carrot root cubes and
Tobacco plant by leaf disc culture 118
Expt. 3 Aseptic organ culture technique, of embryo and ovary 121
Expt. 4 Determine callus growth and calculate the Callusing Index 123
Expt. 5 Study the effect of auxin or cytokinin and the plant growth regulators 126
Expt. 6 Micro-propagation of rice by in vitro organogenesis from embryo 129
Expt. 7 Indirect somatic embryogenesis in carrot (Daucus carrota) 131
Expt. 8 A. In vitro germination of orchid seeds 133
B. In vitro germination of Tobacco, and Tomato seeds 135
Expt. 9 Exact protoplast from plant tissue and demonstrate the somatic cell fusion 137
Expt.10 Production of synthetic seeds and test their viability 140
 Contents ix

5: Biochemistry, Enzyme and Fermentation Technology 145

Expt 1A Estimation of mouse or chicken liver glycogen by anthrone method 145
Expt 1B Preparation of liver homogenate and estimation of liver glycogen 147
Expt 2 Extraction of lipid from egg yolk 148
Expt 3 Separate lipids by thin layer chromatography (TLC) method 149
Expt 4 Protein extraction and estimation from mouse liver 150
Expt 5 Preparation of standard curve of protein (BSA) by Folin-Lowry method and determination
of unknown protein concentration 151
Expt 6 To study the effect of enzyme lactase on milk 153
Expt 7 To immobilize yeast cells by sodium alginate entrapment method and to study the rate
of conversion of glucose by immobilized yeast cells 155
Expt 8 Protease estimation from fruits 157
Expt 9 Extraction of ethyl alcohol from molasses by yeast fermentation 159
Expt 10 Sauerkraut production and to analyze certain parameters during production
(the period of fermentation) 161

6: Cell and Molecular Biology 165

Expt 1 Estimation of nucleic acid. To estimate the RNA isolated from a given sample
OR Spectrophotometric analysis of RNA/ orcinol determination of RNA 165
Expt 2 Estimation of DNA by diphenylamine (DPA) method.
OR Preparation of DNA standard curve through spectrophotometer analysis using
diphenylamine reagent 167
Expt 3 Extraction of Genomic DNA. To extract genomic DNA from a given plant specimen
by using CTAB method (Based on method of Saghai- Maroof et al., 1984) 168
Expt 4 DNA extraction from blood 170
Expt 5 Perform agarose gel electrophoresis for screening the DNA isolated and to estimate
the relative sizes 172
Expt 6 RNA isolation and cDNA preparation.
A. To isolate total RNA from liver tissue by Trizol method 174
B. Complimentary DNA (cDNA) preparation from the isolated total RNA 175
Expt 7 Fractionation of cellular components-
A. Isolation of mitochondria. 176
B. Isolation of chloroplasts from a given plant specimen 177
Expt 8 Separation of proteins by SDS gel electrophoresis 178
Expt 9 Blood smear preparation and screening. To prepare a blood smear and to study
the types of cell present 181
Expt 10 Problems in molecular genetics 183

7: Recombinant DNA Technology 186

Expt 1 Bacterial Transformation. Competent cell preparation. Bacterial transformation and
screening of transformed colonies 186
Expt 2 Isolation of plasmid. Plasmid alkaline lysis-miniprep protocol 188
Expt 3 Detection of plasmid DNA by agarose gel electrophoresis 189
Expt 4 Restriction digestion of DNA and its analysis by agarose gel electrophoresis 191
Expt 5 Polymerase chain reaction—To set up a standard PCR reaction for a specific
gene fragment of interest 193
Expt 6 Gene cloning—To clone a gene of insert in a suitable vector system 194
x Contents

Expt 7 Bacteriophage isolation—To isolate bacteriophage/coliphages from the sewage sample 196
Expt 8 Transformation by co-cultivation techniques in tobacco plants (leaf discs) using Agrobacterium
tumefaciens 197
Expt 9: Medical Biotechnology—To separate blood mononuclear cells by Ficoll hypaque and
observing viable PBMC’s by trypan blue staining 201
Expt 10 Bioinformatics
A. To analyze a given gene sequence for mutation by software analysis 202
B. To analyze a given protein sequence for mutation by software analysis 205

8: Animal Biotechnology and Immunology 207

Expt.1 To learn the basics of animal cell culture 207
Expt. 2 To describe the maintenance of cells in culture 208
Expt. 3 To estimate the number of viable cells using the trypan blue viability test method 210
Expt. 4 Protein extraction and estimation from cell lines by immuno-blotting 211
Expt. 5 Immunology. Bacterial antigen preparation—Preparation of antigens from bacterial
culture routes of inoculation of antigens in rabbit/mice by different routes of immunization 213
Expt 6 Screening for the development or formation of the primary antibody against the inoculated
(administered) antibody by-
A. ‘Haemaglutination’ 215
B. ‘Immuno-diffusion’ methods 215

Annexures 218
Annexure I List of Equipments required for conducting Biotechnology practical 218
Annexure II Common Buffers usually used in Biotechnological experiments 218
Annexure III Stains and Nutrient Media composition for Microbiological experiments 220
Annexure IV Plant Tissue Culture Media Composition 226
Annexure V Answers to the Problems in Chapter II: Genetics and Chapter VI: Cell and
Molecular Biology 228
Annexure VI Chi-square table (χ2 ) 234
Annexure VII References/ Technical Literatures 234
Annexure VIII Addresses of Some Suppliers of Equipments (Minor and Major),
Chemicals, Glass- and Plastic-wares 235
Index 237
 Preface

T
he identification and study of human genes pose one of the greatest challenges to the past and the future
of mankind. In India, two streams of work with genes seems to have come to the forefront, putting Indian
geneticists on the global map. First, a consortium of Asian Scientists, coordinated by the Genome Institute
of Singapore, has analysed variations in genetic sequences from which it may be concluded that all Eastern Asian
population had roots in India. Secondly, the scientists at the Institute of Genomic and Industrial Research claim to
have successfully mapped the human genome sequence for the first time in India, thereby joining, albeit a bit late
in the day, an exclusive club of five nations. The benefits of this achievement are more directly related to the field
of medicine, from predicting diseases in individuals to advanced forms of treatment.
Need for Quality Manpower
This necessitates grooming of personnel from grass-root levels for quality biotechnological research in order to
cope up with the fast-changing scenario. We believe that emphasis on efficient practical applications leads to a
better understanding of theoretical knowledge.
Leaving aside some private research laboratories, a few elite central universities, IITs and NITs, other institutes
across the country need significant up-gradation. Recently, 374 world-class universities have been permitted to set
up campuses countrywide (UGC newsletters) which will create a vacuum for quality biotechnologists [students,
professionals, researchers and scientists], until this gap is bridged by trained hands.
Target Audience: Conceiving the Idea
Keeping in mind the present scenario of practical-class standard in colleges and universities, we conceived the
idea of taking up this project with Tata McGraw Hill Education. This book will be useful for any fundamental
course of Biotechnology, e.g., BSc Biotechnology, BTech Biotechnology and MSc Biotechnology. It will cater to
all major papers of other undergraduate and postgraduate life-science courses like BSc Botany, BSc Zoology, BSc
Biochemistry as well, where Biotechnology is an ancillary subject. Besides, it can be helpful to XI and XII CBSE
students opting for Biotechnology as a subject.
Organisation of the Book: Selection of Practicals
It was done with the hope of emphasising standard work and including cost-effective experiments, so that students
may perform a large number of useful biotechnological practicals. We are aware that there is ample scope for
including innumerable number of experiments, but only those experiments which are of significant importance have
found place in this book. Biotechnological subjects like Genetics, Microbiology, Plant and Animal Tissue Culture,
Biochemistry, Enzyme and Fermentation Technology, Immunology, Cell and Molecular Biology, Recombinant
DNA Techniques, Food, Environment and Medical Biotechnology have been included.
The book consists of eight chapters covering experiments on the above topics, to give an essence in application
of biotechnology compiled in a single book. More than 20 syllabi of different universities, IITs and other renowned
institutions have been scrutinised in selecting these experiments.
xii Preface

The experiments are concisely structured to cover all requisite details. The language is simple and lucid. For
easy understanding, the Table of Contents is segregated into 6 broad sections: Common Laboratory Techniques,
Genetics, Microbiology, Plant Tissue Culture, Cell and Molecular Biology, Recombinant DNA Technology and
Biochemistry.
Besides this, each experiment is explained stepwise with features like
Key terminologies, Aim, Principle/Theoretical Background, Materials required, Examples, Solved Problems,
Step-wise Protocol, Notes, Figures, Data Tabulation—how to record observations, how to draw conclusions,
Precautions, and Frequently Asked viva-voce Questions. Techniques for low-cost methodologies to perform
experiments are also given.
The strong pedagogy includes
Over 50 relevant photographs and illustrations
More than 100 solved and unsolved problems
Over 200 viva-voce questions
8 appendices
Scope and Choice of Experiments
This book will give a wide scope in selecting the practical experiments by teachers and instructors according to
their budget and institutional facilities. It will serve as a readymade reckoner to the teachers, and students can learn
how to conduct and develop a practical step by step. Each of these experiments also includes key terminology and
basic principles underlying them besides possible viva questions.
Acknowledgements
I (Goutam Bhowmik) am indebted to my wife, Dr Sudipa Bhowmik, and daughters Abira and Ruchira for their
constant encouragement and inspiration. Without their support, it would not have been possible to complete this
book. I am also grateful to my student Dr Sujoy Bose, for framing, writing and suggesting constructive ideas
throughout. We would also like to thank Mr Kunal Hore for helping with the illustrations. Also, some reviewers of
the book deserve a special mention for their useful suggestions and comments. Their names are given below.
Partha Roy Indian Institute of Technology (IIT), Roorkee
J Kamalesvari Pachiyapas College of Arts and Science, Chennai
J Joel Gnanadoss Loyola College
Pankaj K Hiradhar Rajasthan University
Feedback
We are also aware of the fact that there may be many shortcomings in the book. Therefore, we without any
prejudice, invite constructive criticisms and useful suggestions from the users. This will also help us to improve
the future editions.
GOUTAM BHOWMIK
SUJOY BOSE
Publisher’s Note
Do you have a feature request? A suggestion? We are always open to new ideas (the best ideas come from you!).
You may send your comments to [email protected] (kindly mention the title and author name
in the subject line).
Piracy-related issues may also be reported.
 Abbreviations
APS Ammonium per sulphate MI mitotic index
aq aqueous min minutes
B5 Gamborg et al. ml milititre
Bis N, N-methylenebisacrylamide mm millimetre
BAP benzyle amino purin mM millimolar
bp base pair MS Murashighe & Skoog
BSA bovine serum albumin MW molecular weight
cm centimeter MSDS material safety data sheet
cM centimorgan ng nano gram
CTAB cetyltrimmethylammonium bromide nm nanometer
2,4-D 2,4 dichlorophenoxy acetic acid NAA 1-napthyl acetic acid
DDH2O double distilled water O.D. optical density
DNA deoxyribonucleic acid PAGE polyacrylamide gel electrophoresis
EDTA ethylenediamine triacetic acid PCR polymerase chain reaction
emf electromotive force PMC pollen mother cell
EtBr ethidium bromide PMSF poly methyl sulfonate
ε molar extinction coefficient pNP p-nitro phenol
FDA flourescein diacetate pNPP p-nitro phenyl phosphate
g gram RBC red blood cells
g/l gram per litre RE restriction enzyme
GA3 gibberellic acid RNA ribonucleic acid
h hour rpm revolution per minute
IAA indole-3- acetic acid RT room temperature
IBA indole-3- butyric acid SDS sodium dodecyl sulfate
kbp kilobase pairs sec second
KIN kinetine SSC saline solution citrate
l litre TE tris-EDTA
LB Luria Bertini TAE tris-acetate-EDTA
lb/in2 pounds per square inch TBE tris-borate-EDTA
mA milliampere TEMED tetramethyethylenediamine
μ micron Tris-Cl tris (hydroxylmethyl) aminomethane
μg/μl microgram per microlitre Chloride
μl microlitre UV ultra violet light
μM micromolar V/V volume per volume
μmol micromoles W/V weight per volume
M molar WBC white blood cells
mg/l milligram per litre xg gravity
mg/ml milligram per millilitre
 1
Introduction to the
Laboratory

1 Biotechnology Biology is the study of living things, which are made of cells. Technology is
about solving problems and making things. Biotechnology uses organisms, cells or parts, to
make and do things we need. These cells may come from microbes, plants or animals.
2 Analytical techniques To know about solving problems critically or make logical reasoning.
3 Application of practical knowledge/experience Use of knowhow or experience of practical for
useful purpose and utility.
4 Laboratory direction Procedures to be maintained or followed in the practical classes.
5 Safety rules Steps to be followed for safeguarding oneself and others.
6 Biohazards Harmful effect which may be caused by microbes or anything which comes from a
living things, viz., animals or plants.
7 Experimental evidence Gathering information indicating whether a proposition is true or valid
after working out a practical or problem or investigation.
8 Interpreting results Find out logical reasons or knowhow for solving any outcome of a
problem.
9 Inquisitiveness Eagerly seeking knowledge or show interest to know something.
10 Laboratory notebook Practical copybook used for recording the findings or results obtained
from doing practical work.
11 Instrumentation The instruments used for measuring some information.

The purpose of this laboratory manual is to help leading to a host of specialties, such as molecular
you learn fundamental laboratory techniques and genetics, microbial physiology, biochemistry,
good laboratory practices that are important in any biochemical engineering, etc., (Fig.1.1). It probes
bioscience laboratory. This course is intended to into the living cells or its constituents to make
introduce you to some of the most widely used products or processes of commercial value. You
experimental procedures in Biotechnology. The will also gain familiarity with some of the types of
development of Biotechnology is dependent on equipment frequently used in biotechnology. The
analytical techniques, and a wide variety of tools biotechnology laboratory course, like all laboratory
and sophisticated equipments. The emergence courses, is an exploration of procedures.
of the science of Biotechnology is due to the Implementation of this manual has been
combined efforts of chemists, physicists, biologists, conceived with an idea that biotechnological
statisticians and engineers. Biotechnology is a concepts depend much on practical guidelines
multidisciplinary field having its roots in the and experiments. Understanding of the theoretical
biological, chemical and engineering sciences concept in biotechnology can come through
2 Analytical Techniques in Biotechnology

protocols and practical knowledge. Quality work ∑ Ensure better understanding of the material
requires thoughtfulness, an ability to recognize by acknowledging the problems related to
problems and a willingness to solve them. These the planned experiments.
are skills and aptitude that are invaluable in any ∑ Read the manual and participate as much as
scientific career. possible in the discussions in order to get full
benefit from the course.
∑ Ask questions in or out of the class. You
should also try to participate in the actual lab
work.
∑ The more effort you put into the course work,
the more you will learn. A practical class is
an opportunity to learn valuable skills.
∑ When in doubt, ALWAYS ASK FOR HELP.

BIOLOGY LAB SAFETY

RULES
For the safety and convenience of everyone
working in the laboratory, it is important that the
biology lab safety rules are observed at all times.
The following rules need to be observed.

(a) Back to the Basics

Fig. 1.1 Multidisciplinary nature of biotechnological science
∑ Place only those materials required for the
day on the benchtop for performing the
BASIC PRE-LABORATORY experiment. All other extra things should be
PREPARATIONS placed in the lab bench storage areas under
the lab benches in order to avoid damage or
∑ Prior planning and conceiving a clear contamination.
theoretical background is essential before ∑ Eating, drinking, applying cosmetics or any
performing any experiment. other hand to mouth activity in the lab should
∑ Go through the instructor’s remarks be avoided.
carefully and be thorough with the practical ∑ Unlabelled material found in the refrigerators,
procedures. incubators or freezers should be destroyed.
∑ Try to understand why a particular practical Always mark the back of the plates with your
experiment has been included and how it is initials, the dates and relevant experimental
related to the subject. data, e.g. strain number.
∑ Analyze its applications and methods to use ∑ Please read the laboratory exercises and
the results in Biotechnology. follow the laboratory directions carefully.
∑ Be sure that all chemicals, glassware, tools Familiarize yourself in advance with the
and equipment are readily available. exercises to be performed.
∑ Make a list of chemicals, reagents, tools, ∑ The biology lab is an important aspect of any
glassware, and equipment needed before you biology course. In order to have a good lab
begin an experiment and check whether these experience, make sure that you follow these
materials are readily available in the lab or biology lab safety rules and any instructions
not. given to you by your lab instructor.
 Introduction to the Laboratory 3

∑ Biology lab safety rules are guidelines designed ∑ Handle all glassware carefully. Notify your
to help keep you safe when experimenting. instructor of any broken glassware. Do not
Some equipment and chemicals in a biology pick up broken glassware with bare hands.
laboratory can cause serious harm. Use the dustpan and brush and dispose of it
∑ It is always wise to follow all lab safety in the bin labelled Broken Glass.
rules. Don’t forget, the most helpful safety ∑ While working with hazardous chemicals,
rule is to use plain common sense. wear gloves, face mask and use safety
∑ The following biology lab safety rules are a glasses or goggles during working with a
sample of the most basic rules that should be trans-illuminator.
followed when in biology lab. Most biology ∑ Work in a way that protects as well as does
labs have the safety rules posted in the lab not disturb your fellow workers and the
and your instructor will most likely go over environment outside the laboratory.
them with you before you begin working. ∑ In case of emergency, immediately contact
∑ Since you will use common facilities, your instructor and report how it happened.
all solutions and everything stored in an ∑ Report all accident and breakages to your
incubator, refrigerator, etc. must be labelled. instructor. If any chemicals come in contact
∑ Before you enter a biology lab, you should with your skin or clothing, first wash the
be prepared for and be knowledgeable about affected area with a large amount of water
any lab exercises that are to be performed. and then report it to your teacher.
That means you should read your lab manual
to know exactly what you will be doing. (d) Handling Chemicals
∑ Review your biology notes and relevant
sections in your biology book before your ∑ The best way for handling chemicals is to
lab begins. Make sure you understand all assume that any chemical you handle is
procedures and purposes, as this will help dangerous. Be sure you understand what
you understand the lab activities you will type of chemicals you are using and how
perform. It will also help you get your they should be handled.
thoughts organized for when you have to ∑ Wear protective eyegear when handling
write your lab report. chemicals.
∑ Always read the label on a reagent bottle ∑ Safety goggles should always be worn when
carefully to make sure it contains the you are working with chemicals or any type
chemical you want. Put the bottle in its
of heating apparatus.
original place immediately after use.
∑ Always handle a flammable liquid with great
(b) Clothing care and keep it away from naked flame.
∑ Some chemicals have the potential to damage ∑ Always handle concentrated acids and alkalis
clothing. Make sure that the clothing you with great care.
wear is something you could do without if it
becomes damaged. As a precaution, wear an (e) Handling Glassware and Equipments
apron or a lab coat. ∑ Be sure you know where to find all safety
∑ Wear proper shoes that can protect your feet equipment in the biology lab. These includes
in case something breaks. Sandals or open- items such as the fire extinguisher, first-aid
toed shoes are not recommended. kit, broken glass receptacles, and chemical
waste bins.
(c) Accident and Injuries ∑ Also be sure to know where the emergency
∑ Report any cut, burn or other injuries to your exits are located and which exit route to take
instructor immediately. in case of an emergency.
4 Analytical Techniques in Biotechnology

(f) Handling Heating Substances 4. Notify your instructor about the spill.
∑ Use the Bunsen burner carefully and with ∑ Place all inoculated material in an assigned
caution. Do not lean over it. place in the lab. Culture tubes should be
stored upright in a stand or a plastic beaker,
∑ Always adjust the Bunsen burner to give a
while petri dishes may be stacked and
luminous flame when not using it (or just
incubated upside down.
simply turn it off).
∑ To avoid contamination and distract
∑ Remember to switch off the electric plug
attention from your experiment, do not use
after you use an electrical heater.
cell phones or other personal media devices
(g) Cleanliness in the laboratory.
∑ Since you have only a limited amount of ∑ If body fluids or bloods are used in a
space to call your own, it is to your advantage laboratory experiment (e.g. blood grouping
to keep your work area clean. experiment), each student should work only
∑ After completing an experiment or when with his or her own sample. All caution
one lab class is over, dispose of all material should be maintained to disinfect the
properly and clean the table top with a punching needle. Remember to dispose of
disinfectant. the used swabs, needles, etc. at the end of
∑ All common areas should be kept free of the experiment in a ‘Biowaste’ container.
culture and dirty glassware, electrophoresis
equipment, etc. should be dealt with (i) Biology Labs Do’s and Don’ts
appropriately. ∑ Don’t run or play in the laboratory
∑ Always clean the oil from the oil immersion ∑ Don’t eat or drink in the lab
lens of the microscope with a piece of lens ∑ Don’t taste any chemicals or substances you
paper or pure silk cloth and xylene. are working with
∑ Return all equipment, reagents and other ∑ Don’t use your mouth for pipetting
supplies to their respective places at the end substances
of the class. ∑ Don’t handle broken glass with bare hands
∑ Always wash and/or sanitize your hands with a ∑ Don’t pour chemicals down the drain without
disinfectant soap before leaving the lab. permission
(h) Handling Microorganisms or Cultures ∑ Don’t operate lab equipment without
permission
∑ Some microorganisms (in microbiology
experiments) used, may be pathogenic, ∑ Don’t perform your own experiments unless
therefore, it is essential to always follow granted permission
proper aseptic techniques while handling ∑ Don’t leave any heated materials unattended
organisms. ∑ Don’t place flammable substances near heat
∑ If you spill a culture, observe the following ∑ Don’t engage in childish antics such as
procedure: horseplay or pranks
1. Immediately throw the culture tube in ∑ Don’t throw regular trash in the ‘Biohazard’
the plastic bin labelled ‘Bio-hazard’, so container.
that no one else touches the contaminated ∑ Never enter the laboratory unless a teacher is
tube. present.
2. After putting off the Bunsen burner, spray
∑ Never remove anything from the laboratory
isopropyl alcohol. After a few minutes,
use paper towels to dry the area. without your teacher’s permission
3. Wash your hands with a disinfectant soap ∑ Never use your bare hands to transfer
and sanitize your hand. chemicals. Use a spatula instead.
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