Laboratory Techniques in Biotechnology
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Questions and Answers

It is advisable to directly smell or taste chemicals to identify them if the label is missing.

False

What are the four main types of chemicals mentioned?

  • Corrosives (correct)
  • Toxic chemicals (correct)
  • Reactive chemicals (correct)
  • Flammables (correct)

    • What precaution should be observed when pouring chemicals?

    Always pour concentrates into the water and not vice-versa

    ______ are chemicals that burn readily and may explode if ignition sources are present.

    <p>Flammables</p> Signup and view all the answers

    What is the purpose of a spectrophotometer?

    <p>To convert light of different wavelengths into a single desired wavelength for analysis.</p> Signup and view all the answers

    What is a common use of a centrifuge?

    <p>All of the above</p> Signup and view all the answers

    A water bath is used to provide variable temperatures for sample incubation.

    <p>False</p> Signup and view all the answers

    An oil bath is a type of heated bath used in a laboratory to heat reaction mixtures. __________ oil is commonly used in modern oil baths.

    <p>Silicone</p> Signup and view all the answers

    What should be worn when washing glassware that contained an irritant or toxic material?

    <p>Eye protection and gloves</p> Signup and view all the answers

    Match the following types of chemicals with their descriptions:

    <p>Toxic agents = Chemicals poisonous to the body and act rapidly Corrosives = Chemicals that can burn, scald, or inflame body tissues Flammables = Chemicals that burn readily and may explode Reactive = Chemicals that require careful handling and may explode violently</p> Signup and view all the answers

    What is the purpose of rinsing glassware three times in tap water, deionized water, and double-distilled water?

    <p>To clean the glassware before drying and storing it in a clean place.</p> Signup and view all the answers

    How should glassware be cleaned if a simple solvent rinse is not sufficient?

    <p>Submerge in a warm concentrated aqueous detergent solution</p> Signup and view all the answers

    Which material is commonly used to make beakers?

    <p>Glass</p> Signup and view all the answers

    A 6M HCl solution is used to soak glassware containing a __________-containing compound.

    <p>metal</p> Signup and view all the answers

    Condensers are used to cool hot vapors in laboratory experiments.

    <p>True</p> Signup and view all the answers

    HF should be used on calibrated volumetrics.

    <p>False</p> Signup and view all the answers

    Erlenmeyer Flask is commonly known as a conical or E-Spot flask and is named after the German chemist Emil __________.

    <p>Erlenmeyer</p> Signup and view all the answers

    Match the following laboratory glassware with their descriptions:

    <p>Burette = Used to dispense known amounts of a liquid reagent Volumetric Flask = Used to measure accurate and precise amount of liquid Graduated Cylinder = Accurately measures the volume of an object Pipette = Transport a measured volume of liquid</p> Signup and view all the answers

    What are the three physical forms of culture media?

    <p>Liquid, semisolid, solid</p> Signup and view all the answers

    Define seed viability.

    <p>Seed viability is a measure of how many seeds are alive and could develop into plants under appropriate conditions.</p> Signup and view all the answers

    Buffer solutions resist changes in _?

    <p>pH</p> Signup and view all the answers

    Buffers must have a pKa value between 6.0 to 8.0.

    <p>True</p> Signup and view all the answers

    What is the aim of preparing 100 ml of 0.1 M acetate buffer of pH 5.2?

    <p>To prepare 100 ml of 0.1 M acetate buffer of pH 5.2.</p> Signup and view all the answers

    What reagents are used in the preparation of acetate buffer?

    <p>Sodium acetate, acetic acid, and water</p> Signup and view all the answers

    Sterilization refers to a process that eliminates or kills all forms of life.

    <p>True</p> Signup and view all the answers

    Sterilization methods include autoclaving, dry-heat, filtration, UV exposure, and ________.

    <p>ethylene oxide</p> Signup and view all the answers

    Match the sterilization method with its description:

    <p>Autoclaving = Uses steam heated to 121–134°C UV Exposure = A radiation method Ethylene Oxide = Gaseous method Dry-Heat = Another thermal method</p> Signup and view all the answers

    What are the two main classifications of plants based on the flowering?

    <p>Cryptogams and Phanerogams</p> Signup and view all the answers

    What is the purpose of surface sterilizing the plant tissue and putting it on a sterile growth medium?

    <p>To prevent contamination by bacteria or fungi</p> Signup and view all the answers

    ___ taxonomy gives a detailed overview of various morphological and anatomical structures of a plant species.

    <p>Plant</p> Signup and view all the answers

    Modern taxonomy of flowering plants includes Palaeontological studies.

    <p>True</p> Signup and view all the answers

    What is the study of detailed pollen grains called?

    <p>Palynology</p> Signup and view all the answers

    Which system of plant classification is based on evolutionary sequence and genetic relationship?

    <p>Phylogenetic systems</p> Signup and view all the answers

    What is the main purpose of a horizontal laminar flow unit?

    <p>To remove bacteria and fungi from the air</p> Signup and view all the answers

    A transfer cabinet provides a sterile environment

    <p>False</p> Signup and view all the answers

    What is the purpose of having a central supply area in the laboratory?

    <p>To have all necessary items readily available for use</p> Signup and view all the answers

    What is the term for the outermost whorl of the flower?

    <p>Calyx</p> Signup and view all the answers

    What is the purpose of taxonomy in botany?

    <p>To characterize, identify, classify, and name plants based on their similarities and differences</p> Signup and view all the answers

    Carl Linnaeus is known as the 'Father of Botany'

    <p>False</p> Signup and view all the answers

    What is the term for the scientific naming system used in taxonomy?

    <p>Binomial nomenclature</p> Signup and view all the answers

    Study Notes

    Laboratory Techniques

    Chemical Use Guideline

    • Always use a product with a label to reference
    • Do not mix chemicals without specific authorization
    • Always use personal protective equipment
    • When pouring chemicals, pour concentrates into the water and not vice versa
    • Never pour chemicals into an empty, unlabeled container
    • Do not store flammable chemicals near a source of heat
    • Pesticides, fungicides, etc. must be stored in a safe and elevated position

    Types of Chemicals

    • Toxic chemicals: poisonous, examples include hydrogen sulfide and cyanide
    • Corrosives: irritants, examples include chlorine and HCl acid
    • Flammables: chemicals that burn readily, examples include gasoline, benzene, and ethyl ether
    • Reactive: require stability and careful handling, examples include nitroglycerine

    Basic Tips of Safe Chemical Handling

    • Read the label
    • Dress properly
    • Follow directions
    • Know emergency procedures
    • Be careful
    • Report any suspected problems
    • Keep your work area neat, clean, and organized
    • Store everything properly

    Basic Rules of Chemical Safety

    • Be aware, be alert, and be alive
    • Don't buy or store chemicals you do not need
    • Store chemicals in their original container
    • Always wear appropriate clothes and work in a safe environment
    • Always dispose of chemicals safely

    Safety Rules of Chemistry Laboratory

    • Protect your eyes and wear appropriate protective clothing
    • Do not apply cosmetics, eat, or drink in the laboratory
    • Pour from large containers to smaller ones (always add acid to water)
    • Work with volatile chemicals under a fume hood
    • Do not smell any chemicals directly and do not pipette solutions by mouth
    • Know the safety of equipment

    Basic Laboratory Procedures

    • Weighing: requires careful weighing of all components
    • Measuring liquids: use calibrated glassware, pipettes, and a hand-operated device
    • Cleaning glassware: follow proper method of cleaning, including soaking and rinsing

    Sterilization

    • Sterilizing glassware and instruments
    • Sterilizing nutrient media
    • Sterilizing plant materials
    • Sterile culture techniques

    Acids and Bases

    • Strong acids: corrosive, examples include hydrochloric (HCl), nitric (HNO3), sulfuric (H2SO4), and hydrobromic (HBr)
    • Weak acids: often organic, examples include formic acid (HCOOH), acetic acid (CH3COOH), salicylic acid (C6H4(OH)COOH), and citric acid (C5H7O5COOH)
    • Bases: ionic compounds containing metal ions and hydroxide ions, examples include sodium hydroxide (NaOH), potassium hydroxide (KOH), calcium hydroxide (Ca(OH)2), magnesium hydroxide (Mg(OH)2), and ammonium hydroxide (NH4OH)

    Salts

    • Ionic compounds composed of metallic ions and nonmetallic ions
    • Salts dissociate in water
    • Salt solutions are generally electrolytes

    pH

    • pH scale measures the hydrogen ion concentration
    • pH 7 indicates a neutral solution, acids are less than 7, bases are greater than 7

    Safety Equipments in Laboratory

    • Eye wash fountain
    • Safety shower
    • Fire extinguisher
    • Emergency exits

    Personal Safety while Handling Pesticides

    • Avoid contact with pesticides
    • Wear all designated safety equipment
    • Be careful of drips and spills
    • Keep hands away from eyes and mouth
    • Wash your hands before smoking, eating, or bathroom breaks
    • Use designated safety equipment according to the WPS statement on the label and regional requirements

    Instruments in Meteorology, Microbiology, and Biochemistry/Chemistry

    • List of instruments includes:
      • Meteorology: sunshine recorder, anemometer, wind vane, pyranometer, Stevenson screen, hygrometer, ordinary rain gauge, self-recording rain gauge, thermometer, and barometer
      • Microbiology: microscope, balance/scale, centrifuge, laminar airflow, spectrophotometer, refrigerator, freezer, autoclave, hot air oven, incubator, pH meter, and water bath
      • Biochemistry/Chemistry: spectrophotometer, balance/scale, centrifuge, stirrer, pH meter, refrigerator, freezer, flame photometer, hot air oven, water bath, EC meter, and atomic absorption spectrophotometer

    Cleaning and Handling Glassware

    • Conventional method of washing glassware: soaking in chromic acid-sulfuric acid bath, followed by rinsing and drying
    • Alternative methods: washing with hot water and commercial detergents, rinsing with distilled and double-distilled water, and drying
    • Cleaning and handling of glassware: inspect, dry, and store in a closed cabinet

    GLASSWARE

    • Beaker: used for stirring, mixing, and heating liquids, available in various sizes
    • Burette: used to dispense known amounts of liquid reagent, measures from the top
    • Condenser: used to cool hot vapors or liquids, maintains a thermal gradient
    • Erlenmeyer/Conical flask: used for titration, has a flat, conical body, and a cylindrical neck
    • Volumetric flask: used to measure a precise amount of liquid, has a flat bottom and long neck
    • Measuring/Graduated cylinder: used to accurately measure the volume of an object, often more accurate than flasks and beakers### Laboratory Equipment and Techniques

    Pipettes

    • Come in various designs for different purposes, with varying levels of accuracy and precision
    • Types of pipettes:
      • Volumetric pipettes (bulb pipettes): used to measure a specific volume of solution with high precision
      • Graduated pipettes: used to measure and dispense small volumes of liquid
      • Micropipettes: used to measure and dispense very small volumes of liquid (less than 1ml)

    Glassware Cleaning

    • Importance of cleaning glassware: essential in chemistry, necessary for accurate results
    • Two levels of cleaning:
      • Quantitative cleaning: required for high-precision applications, such as analytical or physical chemistry
      • Normal cleaning: used for general laboratory purposes
    • General cleaning tips:
      • Disassemble apparatus immediately after use
      • Triple rinse with solvent, followed by tap water and DI water rinse
      • Air dry on a drying rack
    • Specialized cleaning methods:
      • Acidic peroxide solution: used to clean glassware contaminated with organic residues
      • Chromic acid: used to clean glassware, but not recommended due to environmental and health risks
      • Concentrated hydrofluoric acid: used to clean glassware, but not recommended for calibrated volumetrics due to risk of damage

    Microscope

    • Definition: an instrument used to see objects that are too small for the naked eye
    • Types of microscopes:
      • Optical microscope: uses light to image the sample
      • Electron microscope: uses an electron beam to image the sample
    • Rules for using a microscope:
      • Carry the microscope properly
      • Always begin focusing with the 4X objective
      • Use the coarse focus only with the 4X objective in place
      • Use immersion oil only with the 100X objective in place

    Analytical Balance

    • Definition: a class of balance designed to measure small masses in the sub-milligram range
    • Features:
      • Measuring pan inside a transparent enclosure with doors
      • Used to measure accurate masses of chemicals in laboratory experiments

    Laminar Flow Cabinet

    • Definition: a carefully enclosed bench designed to prevent contamination of sensitive materials
    • Features:
      • Air drawn through a HEPA filter and blown in a smooth, laminar flow towards the user
      • Stainless steel construction with no gaps or joints
      • Optional UV-Germicidal lamp for sterilization

    Incubator

    • Definition: a device used to grow and maintain microbiological cultures or cell cultures
    • Features:
      • Maintains optimal temperature, humidity, and other conditions
      • Used in cell biology, microbiology, and molecular biology experiments

    Spectrophotometer

    • Definition: an instrument used to measure the amount of light absorbed by a sample
    • Features:
      • Uses Beer-Lambert's Law
      • Used for analysis of proteins, carbohydrates, and other substances

    Centrifuge

    • Definition: a device used to separate particles or fluids based on density
    • Features:
      • Small table-top centrifuge with maximum revolutions per minute of 6000
      • Used for sediment examination, separation of serum from clotted blood, and concentration of materials

    Magnetic Stirrer

    • Definition: a device used to stir liquids using a rotating magnetic field
    • Features:
      • Used to mix liquids quickly and efficiently

    Water Bath

    • Definition: a water container with an electrically operated heating device
    • Features:
      • Used to incubate samples at a constant temperature over a long period of time
      • Used to warm reagents, melt substrates, or incubate cell cultures
      • Precautions: regular monitoring of water level, disinfectants can be added, and raised to 90°C for decontamination

    Oil Bath

    • Definition: a type of heated bath used to heat reaction mixtures
    • Features:
      • Uses silicone oil or other heat-stable oils
      • Used in conjunction with a hot plate or heating mantle

    Sand Bath

    • Definition: a container filled with heated sand used to provide even heating
    • Features:
      • Used in conjunction with a hot plate or heating mantle
      • Used for chemical reactions and solution preparation

    Fume Hood

    • Definition: a ventilated cabinet used to safely handle and discharge environmentally harmful gases
    • Features:
      • Used for chemical reactions and solution preparation
      • Provides a safe working environment for laboratory personnel### Earthing and Electrical Safety
    • Earthing is a method of protecting against electric shocks by providing a path for fault current to flow to earth.
    • Key function of equipment earthing is to prevent the buildup of static electricity, reducing the risk of electrical discharge in hazardous environments.
    • Current refers to flowing electricity.
    • Earth refers to a connection to the ground.
    • Electrical installation refers to a fixed wiring system.

    Handling of Chemicals and Safety

    • There are four types of chemicals:
      • Toxic agents: poisonous to humans and can act rapidly.
      • Corrosives: irritants that can damage body tissues.
      • Flammables: burn readily and may explode or burn if sparks or flames are present.
      • Reactives: require stability and careful handling, can explode or react violently if not handled properly.
    • Good handling practices:
      • Obtain the minimum amount of chemicals needed.
      • Ensure all containers are clearly labelled.
      • Toxic materials must be locked away.
      • Corrosive substances must be stored securely at a low level.
      • Flammable materials must be stored in specially designed cupboards.
    • Safety precautions:
      • Always wear personal protective equipment (PPE).
      • Never mouth-pipette.
      • Always dilute concentrated acids by adding acid to water.
      • Never carry winchesters by the neck.
      • Always leave benches and balances clean and tidy.
      • Don't mix chemicals without authorization.

    pH Meter and Its Function

    • A pH meter measures the electro-chemical potential between a known liquid and an unknown liquid.
    • The pH meter consists of a glass electrode and a reference electrode.
    • The glass electrode measures the electro-chemical potential of hydrogen ions.
    • To complete the electrical circuit, a reference electrode is needed.
    • The pH meter measures the electrical potential difference between the unknown liquid and the solution inside the glass electrode.
    • Calibration of the pH meter is done using standard solutions with known pH values.

    Preparation of Media and Sterilization

    • Media/microbe culture: a nutrient blend used to support microbial growth.
    • Culture: part of a specimen grown in culture media.
    • Culture media: a medium (liquid or solid) that contains nutrients to grow bacteria in vitro.
    • Types of culture media:
      • Simple (basal, ordinary)
      • Enriched culture media
      • Selective media
      • Differential media (indicators)
    • Properties of media:
      • Supports the growth of bacteria.
      • Nutritive (contains the required amount of nutrients).
      • Suitable pH (neutral to slightly alkaline).
      • Suitable temperature and atmosphere.

    Organic Farming

    • Organic farming: a system that avoids the use of synthetic inputs and relies on crop rotations, crop residues, animal manures, and biological systems of nutrient mobilization and plant protection.
    • Principles of organic farming: care, health, ecology, and fairness.
    • Components of organic farming:
      • Manure
      • Green manuring
      • Vermicompost
      • Bio-fertilizers
      • Crop rotation
      • Animal husbandry
      • Biological management

    Seed Viability

    • Seed viability: a measure of how many seeds are alive and can develop into plants.
    • Why test seed viability: to ensure that seeds stored in a gene bank will grow and produce plants.
    • When to determine viability: at the start of storage and at regular intervals during storage.

    Tissue Culture of Crop Plants

    • Tissue culture: the aseptic culture of plant protoplasts, cells, tissues, or organs under conditions that lead to cell multiplication or regeneration of organs or whole plants.
    • Techniques:
      • Micro propagation
      • Preparation of plant tissue for tissue culture
      • Factors affecting plant tissue culture: growth media, environmental factors, explants source, and genetics.

    Preparation and Standardization of Reagent Solutions

    • Volumetric (titrimetric) analysis: involves determining the volume of a solution of accurately known concentration that is required to react quantitatively with the solution of the substance being determined.
    • Standardization of reagent solutions: essential for accurate analytical results.
    • Types of reagent solutions:
      • Acid-base titration
      • Redox titration
      • Complex formation titration
      • Precipitation titration

    Buffer Solutions

    • Buffer solution: an aqueous solution that resists a change in pH when a small or moderate amount of strong acid or base is added to it.
    • Biological buffers: should meet specific criteria, including a pKa between 6.0 and 8.0, high water solubility, and minimal toxicity towards cells.

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    This quiz covers basic laboratory techniques and safety guidelines in biotechnology, including chemical handling and personal protective equipment usage.

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