Molecular Biology Quiz (Lectures 11 & 12)
This Molecular Biology Quiz covers key techniques such as Southern and Northern blotting, and DNA sequencing. It assesses understanding of molecular genetics, specifically DNA and RNA analysis, and genetic variation through practical applications of these techniques.
- 1.
Southern Blots are used to study...
- A.
DNA only.
- B.
RNA only.
- C.
Proteins only.
- D.
DNA and RNA only.
- E.
DNA and Proteins only.
- F.
RNA and Proteins only.
- G.
DNA, RNA, and Proteins.
Correct Answer
A. DNA only.Explanation
Southern Blots are a laboratory technique used to study DNA. This technique involves the separation of DNA fragments by size using gel electrophoresis, followed by transfer of the DNA fragments to a solid support (such as a membrane). The transferred DNA fragments are then hybridized with labeled DNA probes to detect specific sequences. Therefore, Southern Blots are specifically used to study DNA and not RNA or proteins.Rate this question:
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- 2.
Which is the correct order for steps to be taken in a Southern Blot? I. Detect specific fragments by hybridizing DNA probe (radioactive or non-radioactive) to DNA on membrane. II. Denature dsDNA in gel using NaOH (high pH). III. Digest DNA with restriction enzyme. IV. Visualize hybridized bands with P-imager or film. V. Transfer ("blot") ssDNA from gel to membrane. VI. Size fractionate DNA fragments in agarose gel and stain with EtBr to visualize all DNA.
- A.
III, VI, II, V, I, IV
- B.
II, IV, V, VI, III, I
- C.
IV, I, V, II, VI, III
- D.
III, I, V, VI, II, IV
Correct Answer
A. III, VI, II, V, I, IVExplanation
The correct order for steps to be taken in a Southern Blot is: III, VI, II, V, I, IV. First, DNA is digested with a restriction enzyme (III). Then, the DNA fragments are size fractionated in an agarose gel and stained with EtBr to visualize all DNA (VI). Next, the dsDNA in the gel is denatured using NaOH (II). The ssDNA is then transferred from the gel to a membrane (V). Specific fragments are detected by hybridizing a DNA probe to the DNA on the membrane (I). Finally, the hybridized bands are visualized using a P-imager or film (IV).Rate this question:
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- 3.
What type of blot is used to detect a Restriction Fragment Length Polymorphism (RFLP), in order to analyze genetic variation?
Correct Answer
Southern Blot
Southern Blots
Southern
SouthernsExplanation
A Southern Blot is used to detect Restriction Fragment Length Polymorphisms (RFLPs) and analyze genetic variation. It involves transferring DNA fragments from a gel to a solid support, such as a nitrocellulose membrane, and then hybridizing the DNA with a labeled probe that is complementary to the target sequence. This technique allows for the identification and analysis of genetic variations, such as deletions, insertions, or mutations, within the DNA fragments.Rate this question:
- 4.
Northern Blots are used to study...
- A.
DNA only.
- B.
RNA only.
- C.
Proteins only.
- D.
DNA and RNA only.
- E.
DNA and Proteins only.
- F.
RNA and Proteins only.
- G.
DNA, RNA, and Proteins.
Correct Answer
B. RNA only.Explanation
Northern Blots are used to study RNA only. This technique involves the separation of RNA molecules by gel electrophoresis, followed by transfer onto a membrane and hybridization with a labeled RNA probe. It allows researchers to determine the size and abundance of specific RNA molecules in a sample. Northern Blots are not used to study DNA or proteins, as other techniques like Southern Blots and Western Blots are specifically designed for those purposes.Rate this question:
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- 5.
What are the major differences (besides the specific substance being studied) between northern and Southern blots?
- A.
Northern Blots do not require restriction endonucleases, but the substance being studied must be denatured in heated formaldehyde, in stead of SDS.
- B.
Southern Blots do not require restriction endonucleases, but the substance being studied must be denatured in heated formaldehyde, in stead of SDS.
- C.
Northern Blots require restriction endonucleases, and the substance being studied must be denatured in heated formaldehyde, in stead of SDS.
- D.
Southern Blots require restriction endonucleases, and the substance being studied must be denatured in heated formaldehyde, in stead of SDS.
Correct Answer
A. Northern Blots do not require restriction endonucleases, but the substance being studied must be denatured in heated formaldehyde, in stead of SDS.Explanation
The major difference between northern and Southern blots, besides the specific substance being studied, is that northern blots do not require restriction endonucleases. However, in both northern and Southern blots, the substance being studied must be denatured in heated formaldehyde instead of SDS.Rate this question:
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- 6.
Sanger method of Sequencing DNA description: ________, _____ _____________.
Correct Answer
dideoxy, chain termination
Dideoxy, chain terminationExplanation
The Sanger method of sequencing DNA involves the use of dideoxy nucleotides, which are modified versions of the normal nucleotides. These dideoxy nucleotides lack a 3' hydroxyl group, preventing the addition of further nucleotides during DNA synthesis. This results in the termination of the DNA chain at specific positions, allowing for the determination of the sequence. Therefore, the correct answer is "dideoxy, chain termination."Rate this question:
- 7.
Cycle sequencing is...
- A.
Is an improvement on Sanger method
- B.
Allows automation.
- C.
Includes "454" and "illumina" methods.
Correct Answer(s)
A. Is an improvement on Sanger method
B. Allows automation.Explanation
Cycle sequencing is an improvement on the Sanger method because it utilizes a cyclic reaction to amplify and sequence DNA. This method allows for the automation of the sequencing process, making it more efficient and less labor-intensive. It does not specifically include the "454" and "illumina" methods, but they are examples of sequencing technologies that utilize cycle sequencing principles.Rate this question:
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- 8.
Check all that are true with regards to Sanger Sequencing.
- A.
Syntehsizes new strand of DNA in vitro.
- B.
Includes dideoxy nucleotides (ddNTP) that terminate DNA synthesis.
- C.
Each tube has known dideoxy nt (ddATP, ddGTP, ddTTP, or ddCTP).
Correct Answer(s)
A. Syntehsizes new strand of DNA in vitro.
B. Includes dideoxy nucleotides (ddNTP) that terminate DNA synthesis.
C. Each tube has known dideoxy nt (ddATP, ddGTP, ddTTP, or ddCTP).Explanation
Sanger sequencing is a method used to determine the sequence of DNA. It involves synthesizing a new strand of DNA in vitro, using a DNA template and a primer. The method includes the use of dideoxy nucleotides (ddNTP) that lack a 3' hydroxyl group, which terminates DNA synthesis. Each tube used in Sanger sequencing contains a known dideoxy nucleotide (ddATP, ddGTP, ddTTP, or ddCTP), along with regular nucleotides (dATP, dGTP, dTTP, and dCTP). The termination of DNA synthesis at different points allows the determination of the sequence of the original DNA template.Rate this question:
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- 9.
Differences between Automated Sequencing and Sanger Sequencing:
- A.
DNA labeled with fluorescent dyes instead of (^35)S.
- B.
Dyes are on Dideoxynucleotides(ddNTPs).
- C.
DNA is electrophoresed in polyacrylamide gel.
- D.
DNA labeled with (^35)S instead of fluorescent dyes.
- E.
Dyes are on deoxynucleotides (dNTPs).
- F.
DNA is electrophoresed in agarose gel.
- G.
Detected by exciting dye with laser.
- H.
Not detected by exciting dye with laser.
- I.
Uses one color, and four lanes, one for each nucleotide, for detection.
- J.
Uses one lane, and four colors, one for each nucleotide, for detection.
Correct Answer(s)
A. DNA labeled with fluorescent dyes instead of (^35)S.
B. Dyes are on Dideoxynucleotides(ddNTPs).
C. DNA is electrophoresed in polyacrylamide gel.
G. Detected by exciting dye with laser.
J. Uses one lane, and four colors, one for each nucleotide, for detection.Explanation
The differences between Automated Sequencing and Sanger Sequencing include the use of fluorescent dyes instead of (^35)S for labeling DNA, the presence of dyes on Dideoxynucleotides (ddNTPs) instead of deoxynucleotides (dNTPs), the electrophoresis of DNA in polyacrylamide gel instead of agarose gel, the detection of DNA by exciting the dye with a laser, and the use of one lane and four colors, one for each nucleotide, for detection.Rate this question:
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- 10.
Select those of the following that are true about "Next Generation" sequencing methods:
- A.
"2nd Generation" Sequencing includes "454" Pyrosequencing and Illumina.
- B.
"2nd Generation" Sequencing includes "454" Pyrosequencing only.
- C.
"2nd Generation" Sequencing includes Illumina only.
- D.
"3rd Generation" Sequencing includes "454" Pyrosequencing and Illumina.
- E.
"3rd Generation" Sequencing includes "454" Pyrosequencing only.
- F.
"3rd Generation" Sequencing includes Illumina only.
- G.
"2nd Generation" Sequencing includes single molecule sequencing.
- H.
"3rd Generation" Sequencing includes single molecule sequencing.
Correct Answer(s)
A. "2nd Generation" Sequencing includes "454" Pyrosequencing and Illumina.
H. "3rd Generation" Sequencing includes single molecule sequencing.Explanation
The correct answer is "2nd Generation" Sequencing includes "454" Pyrosequencing and Illumina.,"3rd Generation" Sequencing includes single molecule sequencing. This is because "2nd Generation" sequencing methods refer to the technologies that came after the first-generation Sanger sequencing, and both "454" Pyrosequencing and Illumina are examples of these technologies. On the other hand, "3rd Generation" sequencing methods are the latest advancements in sequencing technology, and single molecule sequencing is one of the key features of these methods.Rate this question:
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- 11.
The main benefit(s) of improvements on sequencing methods is(are)
- A.
Cost by size of molecule being sequenced is lowering.
- B.
Speed of sequencing is rising.
- C.
Cost by size of molecule being sequenced is rising.
- D.
Speed of sequencing is lowering.
Correct Answer(s)
A. Cost by size of molecule being sequenced is lowering.
B. Speed of sequencing is rising.Explanation
Improvements in sequencing methods have led to a decrease in the cost of sequencing larger molecules and an increase in the speed of sequencing. This means that it is now more affordable to sequence larger molecules, making it more accessible for researchers to study complex genomes or conduct large-scale sequencing projects. Additionally, the increase in sequencing speed allows for faster processing of samples, reducing the time required to obtain sequencing results. These advancements in sequencing methods have significantly improved the efficiency and affordability of genetic sequencing.Rate this question:
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- 12.
Which of the following are true about "454" Pyrosequencing?
- A.
It involves a combustion reaction at a molecular level.
- B.
Based on the release of "Pyrophosphate" (P-P) molecule.
- C.
Generates relatively long "reads"(~400bp)
- D.
Generates relatively short "reads"(~40bp)
- E.
Can sequence ~500Mbp DNA/day
- F.
Can sequence ~1000Mbp DNA/day
Correct Answer(s)
B. Based on the release of "Pyrophosphate" (P-P) molecule.
C. Generates relatively long "reads"(~400bp)
E. Can sequence ~500Mbp DNA/dayExplanation
454 Pyrosequencing is a DNA sequencing method that is based on the release of the pyrophosphate (P-P) molecule during the sequencing reaction. This method generates relatively long "reads" of approximately 400 base pairs (bp), allowing for the sequencing of longer DNA fragments. Additionally, it is capable of sequencing approximately 500 million base pairs (Mbp) of DNA per day, making it a high-throughput sequencing technique.Rate this question:
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- 13.
Reporter Genes are:
- A.
"Synthetic" and used to study function of regulatory sequences (promoters, RNA stability, translation regulation, etc.).
- B.
"Natural" and used to study function of regulatory sequences (promoters, RNA stability, translation regulation, etc.).
- C.
Includes "regulatory sequence" from gene of interest.
- D.
Add a coding region for specific "Reporter Protein," one that is the same with an experiment.
- E.
Add a coding region for random "Reporter Protein," one that is the same with an experiment.
- F.
Add a coding region for specific "Reporter Protein," one that is different with an experiment.
- G.
Add a coding region for random "Reporter Protein," one that is different with an experiment.
Correct Answer(s)
A. "Synthetic" and used to study function of regulatory sequences (promoters, RNA stability, translation regulation, etc.).
C. Includes "regulatory sequence" from gene of interest.
D. Add a coding region for specific "Reporter Protein," one that is the same with an experiment.Explanation
Reporter Genes are "Synthetic" and used to study the function of regulatory sequences such as promoters, RNA stability, and translation regulation. They include the "regulatory sequence" from the gene of interest. Additionally, they add a coding region for a specific "Reporter Protein" that is the same as the experiment being conducted. This allows researchers to track and measure the activity of the regulatory sequences in the gene of interest.Rate this question:
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Mar 22, 2023Quiz Edited by
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Mar 12, 2012Quiz Created by
Jimmyd876