Molecular Biology Techniques
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Questions and Answers

What is the primary advantage of microinjection over other molecular techniques?

  • It is a highly efficient method for introducing genetic material into cells.
  • It ensures a high level of expression of the introduced genetic material. (correct)
  • It requires minimal technical expertise and equipment.
  • It allows for the transfer of genetic material into a large number of cells.

    • What is the main function of viral vectors in viral transfection?

  • To replicate the introduced genetic material.
  • To select for cells that have taken up the genetic material.
  • To introduce genetic material into cells as a carrier. (correct)
  • To express the introduced genetic material at high levels.

    • What is the purpose of the 'bla' gene in molecular biology?

  • To express a particular protein at high levels.
  • To glow under near UV light.
  • To allow bacteria to grow in the presence of ampicillin. (correct)
  • To replicate plasmids in bacteria.

    • What is the role of primers in automated DNA sequencing?

    <p>To attach to the template strand and initiate DNA synthesis.</p> Signup and view all the answers

    What is the purpose of terminator bases in automated DNA sequencing?

    <p>To terminate DNA synthesis by attaching to the 3' end of the strand.</p> Signup and view all the answers

    What is the result of gel electrophoresis in automated DNA sequencing?

    <p>Separation of the DNA strands based on size.</p> Signup and view all the answers

    What is the primary advantage of using the GFP gene in molecular biology?

    <p>It allows for the identification of cells that have taken up the genetic material.</p> Signup and view all the answers

    What is the main difference between normal bases and terminator bases in automated DNA sequencing?

    <p>Normal bases have a 3'-OH group, while terminator bases lack a 3'-OH group.</p> Signup and view all the answers

    What is the primary purpose of plating transformed bacteria on a culture medium that includes an antibiotic resistance gene?

    <p>To ensure that only bacteria with the plasmid and antibiotic resistance gene survive</p> Signup and view all the answers

    What is the result of inserting foreign DNA into the MCS part of the LacZ gene?

    <p>The LacZ gene is destroyed, and no blue product is formed in colonies</p> Signup and view all the answers

    What is the primary function of the promoter region of DNA?

    <p>To determine the rate at which mRNA is synthesized</p> Signup and view all the answers

    During plasmid replication, what is replicated along with the bacterial genome?

    <p>The plasmid containing the target gene</p> Signup and view all the answers

    What is the purpose of selective screening in molecular cloning?

    <p>To identify colonies that contain the target gene</p> Signup and view all the answers

    What is the result of the LacZ gene metabolizing X-gal?

    <p>A blue product is formed</p> Signup and view all the answers

    Why is it necessary to allow the bacteria to grow and replicate on the plate during plasmid replication?

    <p>To increase the population of bacteria</p> Signup and view all the answers

    What is the ultimate goal of amplification and screening of a target gene?

    <p>To identify colonies that contain the target gene</p> Signup and view all the answers

    What is the purpose of adding chelating agents like EDTA during DNA preparation?

    <p>To remove divalent cations cofactors</p> Signup and view all the answers

    What is the primary purpose of centrifuging a sample during cell collection?

    <p>To extract cells from the sample</p> Signup and view all the answers

    Which of the following is NOT a common source of DNA?

    <p>Urine samples</p> Signup and view all the answers

    What is the purpose of using a lysis buffer during cell lysis?

    <p>To break down the cell membrane and release DNA</p> Signup and view all the answers

    Why are blood samples collected in EDTA tubes?

    <p>To prevent clotting and preserve DNA</p> Signup and view all the answers

    What is the primary purpose of storing DNA in ethanol at low temperatures?

    <p>To preserve DNA from degradation</p> Signup and view all the answers

    Which type of glial cell is responsible for myelinating cells in the Central Nervous System (CNS)?

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

    Which part of the spinal cord do motor neurons exit through?

    <p>Anterior root</p> Signup and view all the answers

    What is the function of the interneuron in the spinal reflex arc?

    <p>To connect sensory and motor neurons</p> Signup and view all the answers

    What is the effect of the parasympathetic nervous system on heart rate?

    <p>Decreases heart rate</p> Signup and view all the answers

    Which division of the autonomic nervous system is responsible for the 'fight or flight' response?

    <p>Sympathetic nervous system</p> Signup and view all the answers

    What is the function of the neuro-endocrine system?

    <p>To release hormones into the blood</p> Signup and view all the answers

    Which part of the spinal cord gives rise to the lower motor neurons (LMNs) in the parasympathetic nervous system?

    <p>Sacral spinal cord</p> Signup and view all the answers

    What is the effect of the sympathetic nervous system on the digestive tract?

    <p>Decreases peristalsis and motility</p> Signup and view all the answers

    What is the primary function of the Parasympathetic nervous system?

    <p>To regulate unconscious actions, such as heart rate and digestion</p> Signup and view all the answers

    Which type of neuron is responsible for transmitting sensory information from the environment to the CNS?

    <p>Sensory neuron</p> Signup and view all the answers

    What is the main function of Schwann cells in the PNS?

    <p>To form the myelin sheath</p> Signup and view all the answers

    What is the primary function of Astrocytes in the CNS?

    <p>To regulate the blood-brain barrier</p> Signup and view all the answers

    Which type of neuron has only one axon and is responsible for transmitting sensory information?

    <p>Unipolar neuron</p> Signup and view all the answers

    What is the primary function of Ependyma cells in the CNS?

    <p>To control the production and flow of cerebrospinal fluid</p> Signup and view all the answers

    What is the primary function of Microglial cells in the CNS?

    <p>To act as macrophages and clear cellular debris</p> Signup and view all the answers

    How many pairs of cranial nerves are there in the CNS?

    <p>12 pairs</p> Signup and view all the answers

    What is the function of the lateral sulcus?

    <p>Separates the temporal lobe from the frontal and parietal lobes</p> Signup and view all the answers

    What is the primary function of the occipital lobe?

    <p>Visual function</p> Signup and view all the answers

    What is the function of the frontal lobe?

    <p>Short term memory, motor control, higher cognition, personality, foresight, and hindsight</p> Signup and view all the answers

    What is the function of the parietal lobe?

    <p>Processing of tactile and proprioceptive information</p> Signup and view all the answers

    What is the function of the temporal lobe?

    <p>Emotional and visceral responses, learning and memory recall</p> Signup and view all the answers

    What is the central sulcus?

    <p>Divides the frontal and parietal lobe</p> Signup and view all the answers

    What is a gyrus?

    <p>A crest in the brain</p> Signup and view all the answers

    How many pairs of cranial nerves does the CNS have?

    <p>12</p> Signup and view all the answers

    What is the function of the central sulcus in the brain?

    <p>Separates the frontal and parietal lobe</p> Signup and view all the answers

    What is the primary function of the parietal lobe?

    <p>Processing of tactile and proprioceptive information</p> Signup and view all the answers

    What is the function of the lateral sulcus in the brain?

    <p>Separates the temporal lobe from the frontal and parietal lobes</p> Signup and view all the answers

    What is the function of the occipital lobe in the brain?

    <p>Processing of visual information</p> Signup and view all the answers

    What is the function of the temporal lobe in the brain?

    <p>Formation of memory</p> Signup and view all the answers

    How many pairs of cranial nerves are there in the CNS?

    <p>12</p> Signup and view all the answers

    What is the function of the gyrus in the brain?

    <p>A crest in the brain surface</p> Signup and view all the answers

    What is the function of the sulcus in the brain?

    <p>A depression in the brain surface</p> Signup and view all the answers

    What is the result of the failure of the neural tube to close during embryonic development?

    <p>Spina Bifida</p> Signup and view all the answers

    Which of the following reflexes is NOT an approach reflex in infants?

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

    What is the primary function of the ectoderm germ layer during embryonic development?

    <p>Formation of the neural tube</p> Signup and view all the answers

    What is the term for the process by which the neural tube forms during embryonic development?

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

    What is the primary function of the rooting reflex in infants?

    <p>Allows infants to locate and ingest food</p> Signup and view all the answers

    What is the term for the gap in the spine that occurs when the neural tube does not close properly during embryonic development?

    <p>Spina Bifida</p> Signup and view all the answers

    What is the primary function of the Palmar Grasp reflex in infants?

    <p>Enables infants to grasp objects</p> Signup and view all the answers

    What is the result of the failure of the underlying mesoderm and notochord to orchestrate the ectoderm during embryonic development?

    <p>Abnormal neural tube formation</p> Signup and view all the answers

    What is the primary function of the Sympathetic nervous system?

    <p>Stimulating the 'fight or flight' response</p> Signup and view all the answers

    Which type of neuron is responsible for transmitting sensory information from the environment to the CNS?

    <p>Sensory neuron</p> Signup and view all the answers

    What is the primary function of Oligodendrocyte cells in the CNS?

    <p>Forming the myelin sheath in the CNS</p> Signup and view all the answers

    What is the primary function of the Parasympathetic nervous system?

    <p>Promoting digestion and relaxation</p> Signup and view all the answers

    Which type of glial cell is responsible for controlling the production and flow of cerebrospinal fluid?

    <p>Ependyma cells</p> Signup and view all the answers

    What is the primary function of Microglial cells in the CNS?

    <p>Acting as macrophages</p> Signup and view all the answers

    Which type of neuron has only one axon and is responsible for transmitting sensory information?

    <p>Unipolar neuron</p> Signup and view all the answers

    What is the primary function of Astrocyte cells in the CNS?

    <p>Regulating the blood-brain barrier</p> Signup and view all the answers

    Study Notes

    Microinjection

    • Ensures entry and expression into target cells
    • Requires technical expertise and equipment
    • Only relatively few cells can be used

    Viral Transfection

    • Laboratory technique to introduce genetic material into cells using viruses as carriers
    • Process involves the use of viral vectors
    • Does not cause disease

    'Bla' Gene

    • Bacteria with this gene grow in the presence of ampicillin

    GFP Gene

    • Bacteria with this gene glow under near UV light

    Automated DNA Sequencing

    • Laboratory technique to determine DNA nucleotide order
    • Primers attach to the template strand and DNA polymerase reads the template strand
    • Synthesizes a new complementary second strand to match
    • Strand will terminate synthesis when a “Terminator Base” attaches at the 3’ end
    • Terminator bases have dideoxyTTP (ddTTP), which lack 3'-OH group, stopping DNA strand elongation
    • Separate strands by Gel electrophoresis
    • Each terminator base makes up the sequence of DNA, reading from the bottom (5’ to 3’)
    • Identifies the terminal nucleotide in each fragment

    Plasmid Transfer and Drug Resistance

    • Plasmid is transferred from cell to cell, carrying a drug resistance gene
    • Plasmid replication allows the bacteria to grow and replicate on the plate
    • As the bacteria grow, they replicate not only themselves but also the plasmid with gene
    • Population increases
    • Selective screening identifies colonies that likely contain the target gene

    LacZ Selection System

    • The LacZ gene metabolizes X-gal, producing a blue product
    • Insertion of foreign DNA into the MCS part of the LacZ gene destroys the LacZ gene, resulting in no blue product of X-gal formed in colonies (white)

    Promoter Regions

    • Determines the rate at which mRNA is synthesized
    • Amount of protein expressed depends on the choice of promoter

    DNA Preparation and Storage

    • Two ways to prepare and store DNA:
      • Deactivate DNA damaging enzymes by heat/removal of divalent cations cofactors by chelating agents (e.g. EDTA)
      • Storage in ethanol at low temperatures
    • Five popular sources of DNA:
      • Blood samples (must remain unclotted) collected in EDTA tubes
      • Amniocytes (from amniocentesis fluid)
      • Tissue biopsies
      • Cultured cells
      • Paraffin-embedded tissue samples (mainly tumours)

    Cell Collection and Lysis

    • Cells are normally collected using a water mouth wash to dislodge epithelial cells lining the mouth
    • Transfer the sample into a microcentrifuge tube and centrifuge to extract cells
    • During cell lysis, a lysis buffer is used, which consists of various components

    Brain Structure

    • The brain has four lobes: frontal, parietal, temporal, and occipital.
    • The central sulcus divides the frontal and parietal lobes.
    • The lateral sulcus (also called the Sylvian fissure) separates the temporal lobe from the frontal and parietal lobes.
    • A gyrus is a brain crest, and a sulcus is a depression or dip.

    Functions of Brain Lobes

    • Frontal lobe: short-term memory, motor control, higher cognition, personality, foresight, and hindsight.
    • Parietal lobe: processing of tactile and proprioceptive information, comprehension of language, and orientation of individual in space and time.
    • Temporal lobe: auditory, emotional and visceral responses, learning and memory recall, and other functions.
    • Occipital lobe: visual function.

    Neurons and Glial Cells

    • There are three types of neurons: sensory (afferent), relay (interneuron), and motor (efferent).
    • Unipolar neurons have a single axon, bipolar neurons have two axons, and multipolar neurons have multiple axons.
    • Pseudounipolar neurons have a single axon that divides into two branches.
    • Glial cells are neural supporting cells that include Schwann cells, microglial cells, ependyma, astrocyte, and oligodendrocyte.

    Functions of Glial Cells

    • Schwann cells form the myelin sheath in the peripheral nervous system.
    • Astrocyte functions: clearing excess neurotransmitters, regulating the blood-brain barrier, and promoting synapse formation.
    • Microglial cells act like macrophages, clearing cellular debris and dead neurons from nervous tissue through phagocytosis.
    • Ependyma cells control the production and flow of cerebrospinal fluid, brain metabolism, and waste clearance.
    • Oligodendrocyte cells support myelinating cells of the central nervous system.

    Nervous System Components

    • The central nervous system (CNS) has 12 pairs of cranial nerves.
    • The autonomic nervous system has two components: sympathetic and parasympathetic.
    • The somatic motor system includes sensory neurons and motor neurons.

    Spinal Reflex Arc

    • The spinal reflex arc is an involuntary response to a stimulus.
    • The arc involves a sensory neuron, an interneuron, and a motor neuron.
    • The response bypasses the brain, making it faster.

    Autonomic Nervous System

    • The parasympathetic division is responsible for "rest and digest" functions.
    • The parasympathetic division arises from the sacral spinal cord and has various effects on the body, including decreasing heart rate and increasing secretion of tears and saliva.
    • The sympathetic division is responsible for "fight or flight" responses.
    • The sympathetic division arises from the thoracic and lumbar spinal cord and has various effects on the body, including dilating the pupil, increasing heart rate, and constricting blood vessels.

    Neuro-Endocrine System

    • The neuro-endocrine system involves cell bodies in the nucleus of the hypothalamus releasing hormones into the blood by acting on glands.

    The Nervous System

    • The CNS has 12 pairs of cranial nerves.
    • The Autonomic Nervous System has two components: Sympathetic and Parasympathetic.

    Neuron Structure

    • Sensory neurons are afferent.
    • Relay neurons are interneurons.
    • Motor neurons are efferent.
    • Unipolar neuron structure is sensory and has an example.
    • Bipolar neuron structure has an example.
    • Pseudo-unipolar neuron structure has a specific function.
    • Multipolar neuron structure is the most common type.

    Glial Cells

    • Glial cells are neural supporting cells.
    • Examples of glial cells include:
      • Schwann cells
      • Microglial cells
      • Ependyma
      • Astrocyte
      • Oligodendrocyte
    • Astrocyte functions include:
      • Clearing excess neurotransmitters
      • Regulating the blood-brain barrier
      • Promoting synapse formation
    • Microglial cells function as macrophages, clearing cellular debris and dead neurons.

    Brain Structure

    • A gyrus is a brain crest, and a sulcus is a depression.
    • The four lobes of the brain are:
      • Frontal lobe
      • Parietal lobe
      • Temporal lobe
      • Occipital lobe
    • The central sulcus divides the frontal and parietal lobes.
    • The lateral sulcus separates the temporal lobe from the frontal and parietal lobes.

    Lobe Functions

    • Frontal lobe functions include:
      • Short-term memory
      • Motor control
      • Higher cognition
      • Personality
      • Foresight and hindsight
    • Parietal lobe functions include:
      • Processing tactile and proprioceptive information
      • Comprehension of language
      • Orientation of individual in space and time
    • Temporal lobe functions include:
      • Auditory processing
      • Emotional and visceral responses
      • Learning and memory recall
      • Seeking and capturing prey
      • Courtship, mating, and rearing of young
      • Balance between aggressive and communal behavior
      • Formation of memory
      • Homeostasis
    • Occipital lobe function is:
      • Visual function

    Developmental Disorders

    • Anencephaly is a serious birth defect in which a baby is born without parts of the brain and skull.
    • Spinal Bifida is a type of neural tube defect, causing a gap in the spine.

    Reflexes

    • Approach reflexes are present at birth and include:
      • Breathing
      • Rooting
      • Sucking
      • Swallowing
    • Avoidance reflexes include:
      • Coughing
      • Sneezing
      • Blinking
    • Palma grasp is a primitive and involuntary reflex found in infants of humans and most primates.

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    Description

    This quiz covers various techniques used in molecular biology, including microinjection, viral transfection, and gene identification. Test your knowledge of these essential laboratory methods.

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