90 Questions
Who coined the term 'plasmid'?
Joshua Lederberg
What is the main function of plasmids in bacteria?
To carry genes that benefit the survival of the organism
What is the normal number of copies of plasmid found in a single cell called?
Copy number
What is the purpose of phenol-chloroform extraction in DNA purification?
To remove proteins and other contaminants from the DNA
Which of the following is a key application of PCR in forensic science?
Amplifying and sequencing DNA to reveal a person's identity
What is the purpose of annealing in the PCR process?
To bind the primers to the DNA template
What is the typical range for the GC content of PCR primers?
40-60%
What is the purpose of real-time PCR?
To monitor the formation of products as the reaction proceeds
Which detection method in real-time PCR involves the use of a fluorescent reporter molecule and a quencher molecule?
Detection Method 2
What is the purpose of melting curve analysis in real-time PCR?
To assess the dissociation characteristics of double-stranded DNA
Which method involves the synthesis of single-stranded DNA using single-stranded RNA as a template?
Reverse Transcription
Which type of virus uses RNA as its genetic material and replicates through reverse transcription?
Retroviruses
Which enzyme is added to the pyrosequencing reaction mixture to catalyze the reaction of PPi with adenosine 5’ phosphosulfate (APS)?
ATP sulfurylase
What is the limitation of pyrosequencing when it comes to detecting homopolymeric regions?
Homopolymeric regions longer than 10 bases cannot be resolved.
What triggers the release of visible light in pyrosequencing?
The reaction of ATP with luciferin
Which sequencing technology utilizes reversible terminators?
Illumina sequencing
Which of the following is a disadvantage of using qPCR for COVID-19 testing?
Poorer communities cannot afford the qPCR equipment
What is the purpose of a Ct value in real-time PCR?
To determine the number of cycles it took to detect a real signal
What is the purpose of a housekeeping gene in relative quantification of gene expression?
To normalize all Ct values to a baseline experimental condition
How is the fold change in gene expression calculated in qRT-PCR?
Fold change = 2-Δ(ΔCt)
Which of the following is NOT a method for rupturing cellular walls to isolate DNA and RNA from living cells?
Treatment with a hypertonic solution
What is the purpose of centrifugation in the separation of DNA by cesium chloride?
To form a stable, linear gradient of CsCl
Which macromolecule has the highest density and sinks to the bottom of the tube during centrifugation in the separation of DNA by cesium chloride?
RNA
What is the main purpose of extracting nucleic acids from living cells?
To isolate DNA and RNA from living cells
Which of the following is a step involved in RNA-seq library construction?
Fragmentation of RNA
What is the purpose of depleting rRNAs from the total RNA pool in RNA-seq?
To reduce the amount of sequencing reads dominated by rRNAs
Which primer can be used to specifically select for mRNAs in RNA-seq?
Oligo(dT) primer
What is the advantage of using random hexamers for first strand cDNA synthesis in RNA-seq?
Reduces 3' end bias in the sequencing library
What is the role of Ribonuclease H (RNase H) in second strand synthesis in RNA-seq?
Cleaves RNA in RNA:DNA hybrids
What is the purpose of adding an 'A' base to the 3' ends of dsDNAs in RNA-seq library construction?
To enable adapter ligation
Who discovered bacterial conjugation and won the Nobel Prize for it?
Joshua Lederberg
What is the main function of plasmids in bacteria?
To carry genes that benefit the survival of the organism
What is the normal number of copies of plasmid found in a single cell called?
Copy number
What is the purpose of phenol-chloroform extraction in DNA purification?
To remove proteins and impurities from DNA
Which enzyme is responsible for catalyzing the reaction of pyrophosphate (PPi) with adenosine 5’ phosphosulfate (APS) in pyrosequencing?
ATP sulfurylase
What is the limitation of pyrosequencing when it comes to detecting homopolymeric regions?
Homopolymeric regions longer than 10 bases cannot be resolved
Which sequencing technology utilizes reversible terminators?
Illumina sequencing
What triggers the release of visible light in pyrosequencing?
Luciferase
Which detection method in real-time PCR involves the use of two sequence-specific oligonucleotide probes labeled with different dyes?
Hydrolysis probe technique
Which of the following is a key application of PCR in forensic science?
Amplifying and sequencing small amounts of DNA to reveal a person's identity
What is the purpose of melting curve analysis in real-time PCR?
To assess the dissociation characteristics of double-stranded DNA during heating
What is the purpose of annealing in the PCR process?
Allowing the primers to bind to the DNA template
What is reverse transcription?
The synthesis of single-stranded DNA using single-stranded RNA as a template
What triggers the release of visible light in pyrosequencing?
The release of pyrophosphate (PPi) during DNA synthesis
What is the genetic material used by retroviruses?
RNA
What is the purpose of real-time PCR?
To monitor the formation of PCR products as the reaction proceeds
Which of the following is a disadvantage of using Loop-Mediated Isothermal Amplification (LAMP) for COVID-19 testing?
LAMP is prone to false positives
What is the purpose of a cleavable hairpin beacon (CHB) in colorimetric detection of LAMP products?
To increase target specificity
What is the purpose of a standard curve in absolute quantification of gene expression?
To compare results from actual experiments
What is the purpose of normalizing Ct values to a housekeeping gene in relative quantification of gene expression?
To select the correct housekeeping gene
Which of the following is NOT a method for rupturing cellular walls to isolate DNA and RNA from living cells?
Treatment with an isotonic solution
What is the purpose of adding an 'A' base to the 3' ends of dsDNAs in RNA-seq library construction?
To facilitate the ligation of adapters
What is the main function of plasmids in bacteria?
To store genetic information
What is the purpose of melting curve analysis in real-time PCR?
To confirm the specificity of the PCR reaction
Which step is involved in RNA-seq library construction?
Addition of 'A' base to 3' ends
What is the purpose of adding an 'A' base to the 3' ends of dsDNAs in RNA-seq library construction?
To obtain an Illumina library
What is the role of Ribonuclease H (RNase H) in second strand synthesis in RNA-seq?
To degrade only the RNA in RNA:DNA hybrids
Which method involves the synthesis of single-stranded DNA using single-stranded RNA as a template?
Reverse Transcription (RT)
What is the purpose of depleting rRNAs from the total RNA pool in RNA-seq?
To get rid of rRNAs
What is the typical range for the GC content of PCR primers?
30-40%
Which of the following methods can be used to isolate DNA and RNA from living cells?
Treatment with a detergent
How are DNA molecules separated from RNA and protein in the process of cesium chloride centrifugation?
DNA molecules sink to the bottom of the tube
What is the purpose of the polymerase chain reaction (PCR)?
To amplify specific DNA sequences
What is the main function of plasmids in bacteria?
To transfer genetic material between bacteria
Which enzyme is responsible for catalyzing the reaction of pyrophosphate (PPi) with adenosine 5’ phosphosulfate (APS) in pyrosequencing?
ATP sulfurylase
What is the purpose of adding dATP-αS instead of natural dATP in pyrosequencing?
To interfere with the luciferase reaction
What is the limitation of pyrosequencing when it comes to detecting homopolymeric regions?
Homopolymeric regions longer than 10 bases cannot be resolved
What triggers the release of visible light in pyrosequencing?
Luciferase
Which detection method in real-time PCR involves the use of a fluorescent reporter molecule and a quencher molecule?
Detection Method 2
What is the purpose of melting curve analysis in real-time PCR?
To assess the dissociation characteristics of double-stranded DNA during heating
Which type of virus uses RNA as its genetic material and replicates through reverse transcription?
Retrovirus
What is the purpose of reverse transcription in molecular biology?
To generate single-stranded DNA using single-stranded RNA as a template
Which of the following is a key application of qPCR in gene expression analysis?
Comparing the expression levels of different genes
What is the purpose of a standard curve in absolute quantification of gene expression using qPCR?
To determine the amount of targeted nucleic acid in a sample
What is the advantage of using relative quantification in gene expression analysis using qPCR?
It allows for the comparison of expression levels of different genes
What is the formula to calculate the fold change in gene expression using qPCR?
Fold change = 2^(-Δ(ΔCt))
Which of the following is NOT a characteristic of plasmids?
They are responsible for bacterial conjugation
Which of the following is TRUE about the copy number of plasmids?
The copy number of plasmids can vary from one to several hundreds in a single cell
Which of the following is TRUE about CsCl density gradient centrifugation?
CsCl density gradient centrifugation separates plasmid DNA from chromosomal DNA using ethidium bromide
What is the purpose of minipreparation of plasmid DNA?
To isolate plasmid DNA from bacteria
Which step is involved in RNA-seq library construction?
Depletion of rRNAs
What is the purpose of adding an 'A' base to the 3' ends of dsDNAs in RNA-seq library construction?
To facilitate adapter ligation
What is the role of Ribonuclease H (RNase H) in second strand synthesis in RNA-seq?
To degrade RNA in RNA:DNA hybrids
What is the purpose of depleting rRNAs from the total RNA pool in RNA-seq?
To reduce the dominance of rRNAs in sequencing reads
What is the purpose of first strand cDNA synthesis with oligo(dT) in RNA-seq?
To capture an entire transcript
What is the purpose of random hexamers in first strand synthesis in RNA-seq?
To reduce 3' end bias
What is the purpose of real-time PCR?
To monitor the formation of PCR products
What is the typical range for the length of PCR primers?
15 to 30 nucleotides
What is the purpose of annealing in the PCR process?
To allow the primers to bind to the DNA template
What is the purpose of denaturation in the PCR process?
To separate the DNA strands
Test your knowledge on DNA sequencing methods with this quiz! Explore concepts such as RNAse H enzyme, Maxam-Gilbert sequencing, Sanger sequencing, and Pyrosequencing. Learn about the differences between these methods and their principles of operation.
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