Genetics Practical and Lab Safety

Genetics Practical Revision

• Focuses on practical aspects of genetics, including lab safety and procedures.
• Includes information on DNA extraction, lab instruments, and molecular biology techniques.

Safety Rules and Laboratory Instruments

• Emphasizes crucial safety rules in a lab setting.
• Highlights the importance of proper lab practices for preventing accidents, including use of safety equipment and handling of chemicals.
• Introduces different laboratory instruments.

Safety Rules in the Lab

• General lab safety rules include: appearance and attitude, general work procedure, hygiene practice, and emergency procedure.
• Includes specific guidelines like: clean white coat, placing personal items in cabinets, staying in designated areas, listening to the instructor, following instructions, and maintaining proper hygiene.

Appearance and Attitude

• Includes wearing clean white coats.
• Personal items like hair, jewelry, and belongings should be appropriately managed in the lab.
• Proper positioning and respect for colleagues, along with adherence to lab timelines, are emphasized.

General Work Procedures

• Emphasizes meticulous handling of glassware; it should be clean, dry, and unbroken.
• Caution about hazardous chemicals and flames is stressed.
• Proper handling of hot glassware with special tools (holders) is also noted.
• Procedures for heating test tubes (away from the face).
• Handling chemicals, such as avoiding direct sniffing, pouring, mixing, or taste testing.

Hygiene Practice

• Avoiding touching the face, eyes, or clothing with solutions.
• Lab work areas must be cleaned routinely.
• Washing hands thoroughly after any experiments.

Emergency Procedures

• First-aid materials must be readily available.
• Reporting accidents to supervisors promptly.
• Instructions for treating burns (acid and alkali), eye injuries, and fainting are outlined in detail.

Incorrect Actions in Lab

• Never tasting any chemicals.
• Avoiding waving the chemical directly towards the face.
• Don't sniff chemicals directly.
• Never mix chemicals without instructor's permission.

Laboratory Instruments

• Various lab instruments are presented and categorized.
• Instruments for measuring liquid volume, like graduated cylinders, volumetric flasks.
• Instruments for handling chemicals, such as beakers, test tubes, test tube racks, test-tube holders, spatulas, and droppers.
• Tools for specific tasks like heating, stirring, and transferring samples are illustrated.
• Crucibles are introduced for heating certain metals.

Liquid Volume Measurements

• Different types of pipettes suitable for handling various liquid volumes are presented and elaborated upon.
• Automatic pipettes (for accurately measuring small volumes).
• Glass graduated pipettes (for accurate measurement of small volumes).
• Pasteur pipettes (for handling small volumes).
• Graduated cylinders (for measuring larger volumes).
• Volumetric flasks (for precise measurement of volumes and concentrations).

Transfer, Mixing, Boiling, and Storage of Chemicals

• Tools like glass rods for stirring.
• Test tubes and test tube holders for experiments, containing, heating, and mixing.
• Chemical bottles are containers for secure storage of different chemicals and reagents.
• Corrosive substances require specific precautions.

Handling of Chemicals

• Introduce pipette pumps, spatulas, droppers, test tube racks, and test tube brushes.
• Emphasize their respective roles.

DNA Extraction

• Focuses on applying DNA extraction from commercial kits in a practical setting.

DNA Extraction Methods

• The slide provides details on chemical, physical, and combined DNA extraction methods.

Spin Column-Based Nucleic Acid Purification

• Highlights the use of spin column-based nucleic acid purification, a combined method.
• Includes steps like tissue dissociation, cell lysis, DNA binding, wash, and elution.

Spin Columns

• Describe spin columns as small plastic tubes that employ silica resin, selectively binding DNA or RNA based on specific factors.

Principles of DNA Extraction

• Lysis, Precipitation, and Purification are the key steps.

Buccal Mucosa Sample

• Obtain a buccal mucosa sample as the initial step in DNA extraction.

Cell Lysis

• A lysis buffer is used to disrupt the cell membrane within the specimen.
• This action releases all the components of the cell, including DNA.

Centrifugation

• Separates different molecules based on their varying densities.
• Is used for cell collection and DNA precipitation.

Wash steps

• Remove impurities from the DNA sample during extraction using a wash protocol.

Centrifuge (Final Step)

• A centrifuge is used to complete the DNA isolation processes.

Elution Step

• Extracting DNA from the spin column by utilizing a solvent.

Elution Buffer

• Is the key solvent employed to eliminate contaminants from the spin column and release extracted DNA.

Molecular Biology Techniques, Restriction Digest & DNA Electrophoresis

• Introduces advanced molecular biology methods, like restriction digest and DNA electrophoresis. Used for analyzing DNA.

Analyzing DNA

• Outlines the basic steps of analyzing DNA; DNA Extraction, DNA Restriction, and DNA Electrophoresis.

Restriction Enzymes

• Explains restriction enzymes as molecular scissors that cut DNA at specific recognition sites.
• Differentiates sticky ends from blunt ends produced by these enzymes.

DNA Electrophoresis

• Describes the apparatus (components—gel tank, power supply, cover with leads, casting tray, and gel combs).
• Provides diagrams depicting each component.

Parts of Electrophoresis Equipment

• Explains the roles of each piece of electrophoresis equipment. Covers with Leads, Gel Tank, Casting Tray, and Gel Combs.

How to Perform Agarose DNA Gel Electrophoresis

• Explains procedures that lead the process of performing Agarose DNA Gel Electrophoresis

Visualizing DNA by UV Light

• Shows the use of a trans-illuminator and visualization of DNA fragments.

Staining the Sample, Staining the Gel, and DNA Ladder

• Used in gel electrophoresis, for tracking and visualizing DNA fragments.

Molecular Biology Techniques, Blotting Techniques, and DNA Fingerprinting

• Presents blotting techniques and DNA fingerprinting methods.

Blotting Steps

• Demonstrates the different steps in the blotting process, including using an agarose gel, transferring DNA to a membrane, incubating with a radiolabeled probe, visualize DNA bands on a film.

Probe

• Describes probes as a labeled sequence used to detect specific DNA or RNA.

Examples of Labeling Molecules

• Discusses labeling methods: using radioisotopes and nonradioactive molecules with labels like biotin and fluorescent dyes.

Types of Blotting Techniques

• Enumerates various blotting techniques, like Southern blot (DNA), Northern blot (RNA), and Western blot (protein).

DNA Analysis Example

• Illustrates specific application in forensic science/crime investigation.

Introduction to Clinical Bioinformatics (1)

• Introduces clinical bioinformatics, encompassing the usage of computers for collecting, analyzing, and interpreting biological information.

Bioinformatics and Database

• Defines bioinformatics and databases, highlighting their functions.
• Focuses on organized biological data, including genic, proteomic, gene expression, and disease data.

Flowchart for using clinical bioninformatics to analyze the provided case DNA of Osteogenesis imperfecta

• Details a flowchart for using clinical bioinformatics to analyze a specific case.
• Notes steps such as gene prediction, comparing genome, translation of sequence, and determination of protein shape..

Comparison Between Different Databases

• Compares and contrasts different database programs—Blast, Expasy, and Cn3d—with their respective purposes. Describes each program's functions.

This quiz focuses on the practical aspects of genetics, including essential lab safety rules and various laboratory instruments. It covers DNA extraction techniques and outlines crucial safety practices to ensure a secure and effective laboratory environment. Test your understanding of these vital concepts in molecular biology.