Podcast
Questions and Answers
Which enzyme is used to catalyze the formation of a phosphodiester bond between two DNA fragments?
Which enzyme is used to catalyze the formation of a phosphodiester bond between two DNA fragments?
- Polymerase
- Reverse transcriptase
- Endonuclease
- Ligase (correct)
A scientist wants to amplify a specific segment of DNA. Which technique should they use?
A scientist wants to amplify a specific segment of DNA. Which technique should they use?
- Gel electrophoresis
- Southern blotting
- Polymerase Chain Reaction (PCR) (correct)
- Restriction digestion
Which of the following accurately describes the function of endonucleases?
Which of the following accurately describes the function of endonucleases?
- Synthesizing new DNA strands.
- Amplifying segments of DNA.
- Cutting DNA at specific sequences. (correct)
- Joining DNA fragments together.
After cutting a DNA sequence with a specific restriction enzyme, a researcher notices the DNA fragments have overhanging ends. What are these ends called?
After cutting a DNA sequence with a specific restriction enzyme, a researcher notices the DNA fragments have overhanging ends. What are these ends called?
Which of the following is true regarding blunt end ligation?
Which of the following is true regarding blunt end ligation?
What role does reverse transcriptase play in molecular biology?
What role does reverse transcriptase play in molecular biology?
In gel electrophoresis, what property of DNA allows it to be separated?
In gel electrophoresis, what property of DNA allows it to be separated?
During electrophoresis, what causes the separated DNA fragments to move through the gel?
During electrophoresis, what causes the separated DNA fragments to move through the gel?
What is ethidium bromide used for in gel electrophoresis?
What is ethidium bromide used for in gel electrophoresis?
What is the term for a segment of DNA used to locate a specific, complementary sequence?
What is the term for a segment of DNA used to locate a specific, complementary sequence?
What is the primary difference between nuclear DNA and mitochondrial DNA used for identification?
What is the primary difference between nuclear DNA and mitochondrial DNA used for identification?
What are Short Tandem Repeats (STRs)?
What are Short Tandem Repeats (STRs)?
What is required for a individual to be defined as a GMO?
What is required for a individual to be defined as a GMO?
Herbicide-resistant crops were first altered to be uneffected by which herbicide?
Herbicide-resistant crops were first altered to be uneffected by which herbicide?
Which of the following crops produces a natural pesticide?
Which of the following crops produces a natural pesticide?
Which of the following explains Gene Cloning?
Which of the following explains Gene Cloning?
What describes a recombinant plasmid?
What describes a recombinant plasmid?
What main 2 factors are used to form recombinant Plasmids?
What main 2 factors are used to form recombinant Plasmids?
What does bacterial transformation achieve?
What does bacterial transformation achieve?
One way recombinant plasmids into bacterial cells is low, bacteria can be increased through several steps, which of the following is not one of those steps?
One way recombinant plasmids into bacterial cells is low, bacteria can be increased through several steps, which of the following is not one of those steps?
What is indicated when bacterial colonies appear white when grown on X-gal after transformation?
What is indicated when bacterial colonies appear white when grown on X-gal after transformation?
Which of the following is an example of a recombinant protein that can be used to to produce human insluin?
Which of the following is an example of a recombinant protein that can be used to to produce human insluin?
Which phrase describes the end result of Electroporation?
Which phrase describes the end result of Electroporation?
The development of GMOs may involve the silencing of a gene, what does that achieve?
The development of GMOs may involve the silencing of a gene, what does that achieve?
When an engineer is identifying a transgenic organism, that determines that it is transgenic?
When an engineer is identifying a transgenic organism, that determines that it is transgenic?
Golden Rice is known for increased levels of which important vitamin?
Golden Rice is known for increased levels of which important vitamin?
Which vector is responsible used to ingeneer the gox gene in canola plants to create Resistant Roundup canola??
Which vector is responsible used to ingeneer the gox gene in canola plants to create Resistant Roundup canola??
What type of insecticide is the cry1Ac gene?
What type of insecticide is the cry1Ac gene?
CRISPR uses what to precisely change part of a genome by completing actions such as removing or editing sections?
CRISPR uses what to precisely change part of a genome by completing actions such as removing or editing sections?
What did Barrangou and his team identify as the role of CRISPR-Cas9 in bacteria?
What did Barrangou and his team identify as the role of CRISPR-Cas9 in bacteria?
Which component of the CRISPR-Cas9 system acts as a guide, directing the complex to the specific target DNA sequence?
Which component of the CRISPR-Cas9 system acts as a guide, directing the complex to the specific target DNA sequence?
What is the natural function of CRISPR-Cas9 system in bacteria?
What is the natural function of CRISPR-Cas9 system in bacteria?
Before the use of CRSPR or more modern methods, what did early genetic manipulations use to inject cells with?
Before the use of CRSPR or more modern methods, what did early genetic manipulations use to inject cells with?
In the CRISPR-Cas9 system, what role does the PAM (protospacer adjacent motif) sequence play?
In the CRISPR-Cas9 system, what role does the PAM (protospacer adjacent motif) sequence play?
Why is it important to consider whether DNA is circular or linear when determining the number of fragments produced after restriction enzyme digestion?
Why is it important to consider whether DNA is circular or linear when determining the number of fragments produced after restriction enzyme digestion?
What feature of bacterial cells is most critical to their use in recombinant DNA technology for producing human proteins?
What feature of bacterial cells is most critical to their use in recombinant DNA technology for producing human proteins?
In which of the following scenarios would mtDNA analysis be most advantageous over nuclear DNA analysis?
In which of the following scenarios would mtDNA analysis be most advantageous over nuclear DNA analysis?
Flashcards
Polymerases
Polymerases
Enzymes that synthesise DNA.
Ligases
Ligases
Enzymes that join DNA fragments.
Endonucleases
Endonucleases
Enzymes that cut DNA at specific sites.
CRISPR-Cas9
CRISPR-Cas9
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PCR
PCR
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Reverse transcriptase
Reverse transcriptase
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cDNA
cDNA
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Restriction site
Restriction site
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Blunt ends
Blunt ends
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Sticky ends
Sticky ends
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Gel electrophoresis
Gel electrophoresis
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DNA fingerprinting
DNA fingerprinting
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Endonucleases
Endonucleases
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DNA sequencing
DNA sequencing
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Hypervariable regions
Hypervariable regions
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Short tandem repeats (STRs)
Short tandem repeats (STRs)
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Genome editing
Genome editing
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Plasmid
Plasmid
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Antibiotic resistance gene
Antibiotic resistance gene
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Origin of replication (ORI)
Origin of replication (ORI)
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Recombinant plasmids
Recombinant plasmids
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Vector
Vector
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Bacterial transformation
Bacterial transformation
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Electroporation
Electroporation
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Heat shock
Heat shock
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Genetically modified organisms (GMOs)
Genetically modified organisms (GMOs)
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Transgenic organisms
Transgenic organisms
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Study Notes
- Topic 2 covers DNA manipulation techniques and applications, focusing on enzymes used for manipulation, CRISPR-Cas9, PCR, gel electrophoresis, recombinant plasmids, and genetically modified organisms
DNA Manipulation Techniques
- Enzymes consist of polymerase, ligase, and endonucleases, each used to synthesize, join and cut DNA respectively
- CRISPR-Cas9 is a bacterial immune system, adapted to editing an organism's genome by cutting and modifying DNA sequences at precise locations
PCR, Electrophoresis, and Plasmids
- PCR amplifies DNA segments, enabling large quantities of specific sequences to be obtained from very small samples
- Gel electrophoresis sorts DNA fragments by size, primarily used in DNA profiling and forensics
- Recombinant plasmids are vectors used to to transform bacterial cells
Enzymes Manipulating DNA
- Genetic engineers mostly work with genetic material, especially DNA, therefore, the genetic engineer requires tools to cut, join, copy, and separate DNA.
- DNA polymerase was identified by Arthur Kornberg in 1956.
Synthesis of DNA
- If specific genes or sections of DNA are needed, using mRNA as a template is useful
- Reverse transcriptase uses mRNA to build single-stranded DNA with a complementary base sequence
- Second strand created with polymerase
- This DNA is known as cDNA
- Reverse transcriptase has copied the human growth hormone gene
The role of endonucleases
- Endonucleases (restriction enzymes) cleave DNA at restriction sites, with each enzyme recognizing different sequences
- Enzymes can cut at points directly opposite each other, then use blunt ends to ligate
- Other endonucleases cut one strand at one point, then cut the second strand at a point slightly away
- This results in sticky ends, allowing scientists more control over where ligation occurs
Restriction Cutting
- A DNA sample is dissolved then the particular endonuclease is added to it
- The result is the DNA sample will be cut into 2 or more fragments
DNA profiling
- DNA profiling identifies individuals using a technique because each person has unique regions of DNA
- DNA can be amplified through PCR to carry out DNA profiling
- A constant DNA profile enables identification
- DNA can be taken from any cell, blood, somatic cell, etc
- An example of DNA profiling includes comparing one to the national DNA database
Gene editing through CRISPR-Cas9
- CRISPR-Cas9 involves cutting DNA and using a guide to direct the scissors to a site in the DNA
- The guide RNA directs the cellular machinery to a defined genomic target
- Editing means correcting, adding/replacing text, or deleting a sentence
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