Podcast
Questions and Answers
What is the main purpose of an objective lens in a microscope?
What is the main purpose of an objective lens in a microscope?
Which total magnification is achieved when using a high power objective lens with a power of 40X and an ocular lens of 10X?
Which total magnification is achieved when using a high power objective lens with a power of 40X and an ocular lens of 10X?
When is immersion oil necessary in microscopy?
When is immersion oil necessary in microscopy?
What does the field of view in a microscope refer to?
What does the field of view in a microscope refer to?
Signup and view all the answers
What type of microscope is specifically suitable for the observation of large specimen surfaces and 3D information?
What type of microscope is specifically suitable for the observation of large specimen surfaces and 3D information?
Signup and view all the answers
Which of the following statements about lenses in microscopy is true?
Which of the following statements about lenses in microscopy is true?
Signup and view all the answers
What is the primary function of a digital camera when used with a microscope?
What is the primary function of a digital camera when used with a microscope?
Signup and view all the answers
What distinguishes interchromosomal insertion from intrachromosomal insertion?
What distinguishes interchromosomal insertion from intrachromosomal insertion?
Signup and view all the answers
Which of the following describes gene amplification?
Which of the following describes gene amplification?
Signup and view all the answers
What contributes to the identification of mutation hotspots in the genome?
What contributes to the identification of mutation hotspots in the genome?
Signup and view all the answers
How do mitochondrial DNA mutations affect cellular function?
How do mitochondrial DNA mutations affect cellular function?
Signup and view all the answers
What is a translocation in genetic terms?
What is a translocation in genetic terms?
Signup and view all the answers
What constitutes a molecule?
What constitutes a molecule?
Signup and view all the answers
Which of the following elements make up over 90% of the human body?
Which of the following elements make up over 90% of the human body?
Signup and view all the answers
What is the main characteristic of ionic bonds?
What is the main characteristic of ionic bonds?
Signup and view all the answers
How do double covalent bonds differ from single covalent bonds?
How do double covalent bonds differ from single covalent bonds?
Signup and view all the answers
What defines a polar molecule, such as water?
What defines a polar molecule, such as water?
Signup and view all the answers
Which statement about covalent bonding is true?
Which statement about covalent bonding is true?
Signup and view all the answers
Which of the following best describes the role of buffers in biological systems?
Which of the following best describes the role of buffers in biological systems?
Signup and view all the answers
What is an isotope?
What is an isotope?
Signup and view all the answers
What primarily makes organic molecules different from other types of molecules?
What primarily makes organic molecules different from other types of molecules?
Signup and view all the answers
What is the advantage of artificial gene synthesis compared to traditional methods?
What is the advantage of artificial gene synthesis compared to traditional methods?
Signup and view all the answers
What is the function of a microarray in genetic research?
What is the function of a microarray in genetic research?
Signup and view all the answers
Which factor can lead to spontaneous mutations?
Which factor can lead to spontaneous mutations?
Signup and view all the answers
What are germline mutations?
What are germline mutations?
Signup and view all the answers
Which of the following statements about mutations is correct?
Which of the following statements about mutations is correct?
Signup and view all the answers
What is the primary cause of induced mutations?
What is the primary cause of induced mutations?
Signup and view all the answers
What role do chemical mutagens play in DNA?
What role do chemical mutagens play in DNA?
Signup and view all the answers
How does the principle of DNA microarrays work?
How does the principle of DNA microarrays work?
Signup and view all the answers
In the context of DNA, what are purine and pyrimidine bases known for?
In the context of DNA, what are purine and pyrimidine bases known for?
Signup and view all the answers
What is the significance of clustering expression profiles in research?
What is the significance of clustering expression profiles in research?
Signup and view all the answers
What is the primary purpose of DNA cloning?
What is the primary purpose of DNA cloning?
Signup and view all the answers
Which of the following best describes plasmids?
Which of the following best describes plasmids?
Signup and view all the answers
What technique uses DNA polymerase to amplify specific DNA sequences?
What technique uses DNA polymerase to amplify specific DNA sequences?
Signup and view all the answers
Which of the following vectors is often used for cloning in E. coli?
Which of the following vectors is often used for cloning in E. coli?
Signup and view all the answers
What is not a common type of molecular hybridization technique?
What is not a common type of molecular hybridization technique?
Signup and view all the answers
Which method is used for introducing mutations into DNA?
Which method is used for introducing mutations into DNA?
Signup and view all the answers
What are the types of blotting used in molecular biology for different purposes?
What are the types of blotting used in molecular biology for different purposes?
Signup and view all the answers
What is the primary role of a vector in DNA cloning?
What is the primary role of a vector in DNA cloning?
Signup and view all the answers
Which of the following statements is false regarding CRISPR/Cas9?
Which of the following statements is false regarding CRISPR/Cas9?
Signup and view all the answers
Which of the following methods is not a function of gel electrophoresis?
Which of the following methods is not a function of gel electrophoresis?
Signup and view all the answers
Which of the following best describes the structure of smooth muscle fibers?
Which of the following best describes the structure of smooth muscle fibers?
Signup and view all the answers
What is a distinguishing feature of cardiac muscle tissue?
What is a distinguishing feature of cardiac muscle tissue?
Signup and view all the answers
Which of the following functions is NOT performed by skeletal muscles?
Which of the following functions is NOT performed by skeletal muscles?
Signup and view all the answers
How do skeletal muscle fibers differ from smooth muscle fibers?
How do skeletal muscle fibers differ from smooth muscle fibers?
Signup and view all the answers
What role do the neuromuscular junctions play in muscle contraction?
What role do the neuromuscular junctions play in muscle contraction?
Signup and view all the answers
What is one key characteristic of the sliding filament model of muscle contraction?
What is one key characteristic of the sliding filament model of muscle contraction?
Signup and view all the answers
Which type of muscle tissue displays rhythmic contractions without nervous stimulation?
Which type of muscle tissue displays rhythmic contractions without nervous stimulation?
Signup and view all the answers
What is the term for the muscle that is contracting during movement?
What is the term for the muscle that is contracting during movement?
Signup and view all the answers
Which of the following is NOT a component of a muscle fiber?
Which of the following is NOT a component of a muscle fiber?
Signup and view all the answers
What type of myofilament is composed of myosin?
What type of myofilament is composed of myosin?
Signup and view all the answers
Which structure penetrates the muscle fibers and comes close to the sarcoplasmic reticulum?
Which structure penetrates the muscle fibers and comes close to the sarcoplasmic reticulum?
Signup and view all the answers
What is the function of myoglobin in muscle fibers?
What is the function of myoglobin in muscle fibers?
Signup and view all the answers
What would happen if both the agonist and antagonist muscles contract simultaneously?
What would happen if both the agonist and antagonist muscles contract simultaneously?
Signup and view all the answers
Which part of a muscle fiber is responsible for calcium storage?
Which part of a muscle fiber is responsible for calcium storage?
Signup and view all the answers
Where are myofibrils located within a muscle fiber?
Where are myofibrils located within a muscle fiber?
Signup and view all the answers
What provides energy for muscle contraction?
What provides energy for muscle contraction?
Signup and view all the answers
What is the role of myoglobin in muscle fibers?
What is the role of myoglobin in muscle fibers?
Signup and view all the answers
What is the structure that separates sarcomeres in muscle fibers?
What is the structure that separates sarcomeres in muscle fibers?
Signup and view all the answers
What happens to the I band during the contraction of a muscle fiber?
What happens to the I band during the contraction of a muscle fiber?
Signup and view all the answers
Which of the following correctly describes thick myofilaments?
Which of the following correctly describes thick myofilaments?
Signup and view all the answers
What is the function of the sarcoplasmic reticulum in muscle fibers?
What is the function of the sarcoplasmic reticulum in muscle fibers?
Signup and view all the answers
Which statement is true regarding the sliding filament model of muscle contraction?
Which statement is true regarding the sliding filament model of muscle contraction?
Signup and view all the answers
What primary component gives rise to muscle striations?
What primary component gives rise to muscle striations?
Signup and view all the answers
What structure extends into the muscle fiber and conducts impulses to trigger calcium release?
What structure extends into the muscle fiber and conducts impulses to trigger calcium release?
Signup and view all the answers
Which region of the sarcomere contains only thick myofilaments?
Which region of the sarcomere contains only thick myofilaments?
Signup and view all the answers
What mechanism describes the continuous sliding action of myosin and actin filaments during muscle contraction?
What mechanism describes the continuous sliding action of myosin and actin filaments during muscle contraction?
Signup and view all the answers
Which of the following sources of energy for muscle contraction is stored directly in muscle cells?
Which of the following sources of energy for muscle contraction is stored directly in muscle cells?
Signup and view all the answers
Which pathway for ATP production in muscle cells requires oxygen?
Which pathway for ATP production in muscle cells requires oxygen?
Signup and view all the answers
What role do calcium ions (Ca2+) play in muscle contraction?
What role do calcium ions (Ca2+) play in muscle contraction?
Signup and view all the answers
As the duration of exercise increases, which source of energy is predominantly utilized by muscles?
As the duration of exercise increases, which source of energy is predominantly utilized by muscles?
Signup and view all the answers
What action occurs when acetylcholine (ACh) binds to receptors in the sarcolemma?
What action occurs when acetylcholine (ACh) binds to receptors in the sarcolemma?
Signup and view all the answers
What is the primary role of calcium ions (Ca2+) in muscle contraction?
What is the primary role of calcium ions (Ca2+) in muscle contraction?
Signup and view all the answers
What sequence of events occurs first in the sliding filament theory of muscle contraction?
What sequence of events occurs first in the sliding filament theory of muscle contraction?
Signup and view all the answers
What happens after the power stroke in muscle contraction?
What happens after the power stroke in muscle contraction?
Signup and view all the answers
Which component is crucial for the removal of cross-bridges during muscle contraction?
Which component is crucial for the removal of cross-bridges during muscle contraction?
Signup and view all the answers
What is found in the synaptic vesicles at an axon terminal?
What is found in the synaptic vesicles at an axon terminal?
Signup and view all the answers
Which area describes the junction between a motor neuron and a muscle fiber?
Which area describes the junction between a motor neuron and a muscle fiber?
Signup and view all the answers
How does tropomyosin affect muscle contraction?
How does tropomyosin affect muscle contraction?
Signup and view all the answers
What role does the axon play in muscle fiber contraction?
What role does the axon play in muscle fiber contraction?
Signup and view all the answers
What occurs when the nerve signal reaches an axon terminal?
What occurs when the nerve signal reaches an axon terminal?
Signup and view all the answers
Study Notes
Biotechniques (BMS 34010A)
- Course offered during Fall semester 2023-2024
- Instructor: Dr. Tania Tahtouh
- Email: [email protected]
Basic Principles: Biosafety in the Laboratory
- Laboratory settings present potential hazards.
- Safety procedures protect personnel and equipment.
- Disposal of contaminated materials/chemicals follows strict protocols.
Potential Laboratory Hazards
- Corrosive chemicals, sharp tools/glass, and open flames pose risks.
- Bunsen burners require extreme caution.
- Infectious organisms also pose hazards, classified into biosafety levels (BSL 1-4).
- Most undergrad labs use BSL 1 organisms.
Standard Practices in the Laboratory
- Lab coats are essential for protection against contamination.
- Lab coats should be long and not restrict movement.
- Lab coats are not chemical protection suits.
- Gloves should not touch surfaces outside the lab.
- Eating, drinking, chewing gum, or pipetting by mouth are forbidden in the lab.
- No makeup or contact lenses adjustments in the lab.
- Work area must be clean with only necessary materials.
- Cell phones, bags, and other items should be stored away from workbench.
- Avoid leaving active experiments unattended.
- Properly disinfect the workbench before and after each experiment.
- Maintain clean hands and use correct hand washing techniques.
- Unauthorized experiments are forbidden.
- Long hair must be tied up, and loose clothes must be secured when working with open flames.
- Safety glasses or face shields required when working with UV light or chemicals.
- Closed-toe shoes are mandatory to avoid chemical spills and sharp objects.
- Know location of safety equipment: eyewash, chemical shower, first-aid kit, and fire extinguisher.
- Follow chemical hygiene plans and use fume hoods/biosafety cabinets for chemical handling.
- Report spills, accidents, or injuries immediately.
Biohazard Bin
- Biohazardous waste goes in appropriately labeled bins/bags.
- Non-reusable items with contact to infectious material, body fluids, or animal tissue.
- Handled gloves must also be disposed of.
- Items can be autoclaved before disposal.
Sharps Containers & Trash Cans
- Sharps containers for broken glass, needles, used scalpels, glass pipettes, and other sharp objects.
- Non-contaminated items (paper, etc.) go in regular trash cans.
Reusable Materials
- Materials needing sterilization: autoclave after use (e.g., culture tubes and instruments).
- Materials not needing sterilization: wash in sink after use (e.g., dissection tools and glassware).
General Rules for Handling Chemicals
- Keep containers closed.
- Never handle wrong or unmarked reagents.
- Avoid carrying bottles by the neck or against the body.
- Labels should face up when pouring.
- Only place indicated chemical into bottle.
- Handle concentrated acids with caution (add acid to water and stir continuously).
- Disposal of some chemicals requires special procedures.
The Importance of Hand Washing
- Hand washing is crucial in labs, especially microbiology.
- Hands can come into contact with infectious agents, chemicals, stains, and other hazardous materials.
- Wash hands before, after, and whenever contact with potential hazards occurs.
Steps for Correct Hand Washing Technique
- Remove all jewelry.
- Wet hands and apply soap.
- Rub hands together to make a lather.
- Scrub for 20 seconds (e.g., "Happy Birthday" twice).
- Rinse and dry hands with a clean towel/air dry.
Metric Systems
- The SI system (International System of Units) is the standard used by scientists.
- The United States is the only industrialized nation to continue using the English system.
- SI system is based on the number 10.
- The SI system includes measurement units for length, mass, volume, temperature, and time.
Microscopy: Types and Principles
-
Biological specimens often require microscopic visualization.
-
Various types of microscopes exist for different applications.
-
Types of microscopes include Light (compound, binocular, phase contrast, fluorescent, confocal) and Electron (transmission, scanning).
-
Learning Outcomes: Recognize principles/uses of different microscopes, parts of a compound light microscope, total magnification, microscope rules of use, focus methods, field of view, longitudinal/cross sections, and resolution.
-
Light Microscopes: Simplest consists of a single glass lens. -Specimen prep minimal. -Focusing by moving specimen relative to lens.
-
Compound Light Microscopes: Multiple lenses (condenser, objective, eyepiece).
-
Parts of Compound Light Microscope
- Light source (lamp).
- Condenser.
- Objectives.
- Eyepiece.
- Stage & Clips.
- Diaphragm
- Coarse & Fine Focus Knobs.
-Lens types and Magnification.
-
Field of view (at low and high power).
-
Longitudinal & cross sections.
-
Resolution.
-
Scanning and Transmission Electronic Microscopes: Greater magnification to view smaller objects
-
Stereomicroscope: Used to observe large specimens' surfaces in 3-D.
-
Phase contrast microscopy: Improves contrast for transparent and colorless specimens.
-
Fluorescent microscopy: High contrast for specific labeling of cellular components.
-
Fluorescents and stains (RFP, GFP, and DAPI).
-
Immunofluorescence: Using antibodies labeled with fluorescent dyes.
-
Confocal microscopy: Produces optical sections of thick specimens.
-
Microscope use rules outlined in slide deck
DNA Cloning Overview
- Inserting the target DNA into the plasmid vector.
Cut and Paste DNA: Plasmids
- Circular
- Self-replicating.
- Host cell/chromosome separate.
Cut and Paste DNA: Euk expression vectors
-Eukaryotic DNA containing virus restriction sites and/or terminal repeats.
Cut and Paste DNA: Restriction
- Restriction Sites: Specific 4-8 bp.
- Type II Restriction Enzymes: Highly-specific, blunt, or sticky ends.
Cut and Paste DNA: Restriction (Inserting DNA sample into plasmid):
- Enzymes cut DNA at restriction sites, and ligase joins the fragments.
DNA Transfer into Cells (Transformation, Transduction, Transfection)
- Transformation (prokaryotic) - Non-viral method
- Transduction (prokaryotic/eukaryotic) - Viral method
- Transfection (eukaryotic) - Non-viral method
Chromosome Integration
- Integrase splices viral DNA into the cellular chromosome.
Cellular Screening
- Vectors derived from natural plasmids; genetically modified for convenient use, some more advanced than others in structure .
DNA Extraction
- Lysis
- Precipitation
- Purification
- Elution
Extraction of Plasmids
- Separation of plasmid/chromosomal DNA through differential denaturation and neutralization.
DNA Cloning Overview
- Gene amplification and digestion/ligation.
- Transformation & plasmid isolation/sequencing.
Crispr/Cas9
- A genome editing technique in bacteria.
- Creation of specific double-strand breaks at the target locus.
- Types: NHEJ and HR
DNA Synthesis
- Artificial gene synthesis not requiring DNA template.
DNA Array (Microarray)
- Tool for detecting gene expression.
Mutations and Causes
- Spontaneous Mutations: Errors in replication or chemical instability of DNA bases.
- Induced Mutations: Exposure to environmental factors.
Main Types of Mutations
-
Germline: Inherited, in gametes.
-
Somatic: In other body cells.
-
Point mutations: Substitutions, insertions, or deletions of single bases.
-
Chromosomal mutations: Deletions. Duplications. Inversions, insertions, translocations.
-
Copy Number Variation (CNV): Changes in the number of tandem copies of a locus.
Point Mutations
- Missense: Different amino acid.
- Nonsense: Premature stop codon.
- Synonymous: Same amino acid.
- Neutral: No effect.
Point Mutations: Categorization
- Loss-of-function: Decreased/absent gene product.
- Gain-of-function: Increased gene product or new activity
Chromosomal Mutations
- Deletion: Loss of chromosome region.
- Duplication: Repetition of chromosome region.
- Inversion: Flipping and reinserting a chromosome region.
- Insertion: Moving a chromosome region to another.
- Translocation: Rearrangement of chromosome regions.
Copy Number Variation (CNV)
- Gene amplification: Increasing tandem copies of a locus.
- Expanding trinucleotide repeat: Expanding repeated sequences.
Mutation Hotspots
- Regions with higher recombination rates or accessibility to mutagens.
- Repetitive sequences are often hotspots.
Mitochondrial DNA Mutations
-
Inherited changes can cause growth/development/function problems.
-
mtDNA mutations affect mitochondria's energy generation capacity.
-
References: Provide links to relevant articles & resources.
Studying That Suits You
Use AI to generate personalized quizzes and flashcards to suit your learning preferences.
Description
This quiz covers the essential biosafety principles in laboratory settings, focusing on the identification of potential hazards and standard practices to ensure safety. It is designed for students enrolled in the Biotechniques course (BMS 34010A) for the Fall semester of 2023-2024.
