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Gel Electrophoresis in Nucleic Acid Analysis
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Gel Electrophoresis in Nucleic Acid Analysis

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Questions and Answers

What is the primary function of the CRISPR-associated protein (Cas) enzyme?

  • To recognize folded nucleic acids
  • To cleave the invading DNA at specific locations (correct)
  • To guide the Cas enzyme to the matching invading DNA
  • To transcribe short RNA sequences from the CRISPR spacer regions
  • What is the main advantage of CRISPR/Cas9 over traditional restriction enzymes?

  • CRISPR/Cas9 can recognize folded nucleic acids
  • CRISPR/Cas9 can only be used for DNA analysis
  • CRISPR/Cas9 can only cleave single-stranded nucleic acids
  • CRISPR/Cas9 provides the specificity of restriction enzymes with the versatility of guiding cuts to any sequence site (correct)
  • What is the primary purpose of the Southern blot?

  • To analyze RNA structure and expression
  • To analyze specific DNA regions in a complex genome by RFLP (correct)
  • To screen for mutations and polymorphisms
  • To detect proteins and protein modifications
  • What is the primary goal of genomics?

    <p>To analyze hundreds to thousands of targets or whole genomes</p> Signup and view all the answers

    What is the purpose of the spacer sequences in the CRISPR system?

    <p>To serve as adaptive immunity with memory of the invading DNA</p> Signup and view all the answers

    What is the primary application of CRISPR/Cas9 in the laboratory?

    <p>To alter DNA at user-defined locations</p> Signup and view all the answers

    What is the main advantage of CRISPR RNA over traditional restriction enzymes?

    <p>CRISPR RNA can lead transcription activators, repressors, gene promoters, or reporter molecules to target sequences</p> Signup and view all the answers

    What is the purpose of a nucleating agent in liquid acrylamide solutions?

    <p>To facilitate the solidification of acrylamide</p> Signup and view all the answers

    What is the primary function of a density agent in a loading solution?

    <p>To facilitate the loading of the sample into the wells</p> Signup and view all the answers

    What is the purpose of a fluorescent stain in agarose gel electrophoresis?

    <p>To visualize the nucleic acid fragments</p> Signup and view all the answers

    What is the primary advantage of capillary electrophoresis over agarose gel electrophoresis?

    <p>Increased sensitivity</p> Signup and view all the answers

    What is the primary purpose of comparative genomic hybridization arrays?

    <p>To detect amplifications or deletions in DNA</p> Signup and view all the answers

    What type of array is used to measure gene expression levels?

    <p>RNA expression arrays</p> Signup and view all the answers

    What is the function of a platinum wire in agarose gel electrophoresis?

    <p>To establish a current through the gel</p> Signup and view all the answers

    What is the term for the process of detecting chromosome microdeletions using microarrays?

    <p>Virtual karyotyping</p> Signup and view all the answers

    What is the purpose of a molecular weight standard in agarose gel electrophoresis?

    <p>To estimate the size of the DNA fragments</p> Signup and view all the answers

    What is the primary difference between agarose gel electrophoresis and capillary electrophoresis?

    <p>The presence of a gel matrix</p> Signup and view all the answers

    What is the purpose of labeling the sample material in microarray analysis?

    <p>To allow for hybridization of the sample to the probes</p> Signup and view all the answers

    What is the purpose of a laser in capillary electrophoresis?

    <p>To excite the fluorescent labels</p> Signup and view all the answers

    What is the term for the type of array that can be used to determine DNA nucleotide sequence?

    <p>High-density oligonucleotide arrays</p> Signup and view all the answers

    What is the purpose of using a reference material in microarray analysis?

    <p>To compare the levels of gene expression between the test and reference samples</p> Signup and view all the answers

    What is the advantage of using multiple capillaries in capillary electrophoresis?

    <p>Ability to analyze multiple samples simultaneously</p> Signup and view all the answers

    What is the purpose of a visual dye in a loading solution?

    <p>To visualize the sample during electrophoresis</p> Signup and view all the answers

    What is the term for the type of array that uses beads as the solid support?

    <p>Bead array</p> Signup and view all the answers

    What is the purpose of using fluorescent labels in bead array assays?

    <p>To detect the presence of the target protein or nucleic acid</p> Signup and view all the answers

    What is the advantage of bead array assays over other types of arrays?

    <p>They can detect multiple targets simultaneously</p> Signup and view all the answers

    What is a clinical application of bead array assays?

    <p>HLA typing</p> Signup and view all the answers

    What is the primary function of restriction enzymes in DNA analysis?

    <p>To separate phosphodiester bonds between nucleotides</p> Signup and view all the answers

    What is the purpose of molecular weight standards in capillary gel electrophoresis?

    <p>To enable instrument to automatically assess fragment size</p> Signup and view all the answers

    What is the primary mechanism of DNA separation in capillary gel electrophoresis?

    <p>Electrokinetic process</p> Signup and view all the answers

    What is the function of CRISPR-Cas9 system in DNA analysis?

    <p>To recognize and bind to specific nucleotide sequences</p> Signup and view all the answers

    What is the purpose of formamide in preparing DNA samples for capillary gel electrophoresis?

    <p>To denature DNA into single strands</p> Signup and view all the answers

    What is the result of DNA digestion with restriction enzymes?

    <p>Production of DNA fragments of varying sizes</p> Signup and view all the answers

    What is the characteristic of DNA fragments produced by restriction enzymes?

    <p>Fragments are specific to the DNA sequence</p> Signup and view all the answers

    What is the purpose of nucleic acid tests?

    <p>To detect changes in DNA sequence</p> Signup and view all the answers

    What is the name of the first restriction enzyme isolated from E. coli?

    <p>EcoR1</p> Signup and view all the answers

    What is the primary application of restriction fragment length polymorphism (RFLP) analysis?

    <p>All of the above</p> Signup and view all the answers

    What is the primary purpose of electrophoresis in the analysis of nucleic acids?

    <p>To observe the sizes or amounts of nucleic acid</p> Signup and view all the answers

    Which type of gel is more suitable for separating very large nucleic acids of tens of thousands of base pairs?

    <p>Agarose gel at low concentrations</p> Signup and view all the answers

    Why do smaller nucleic acid chains move faster through the gel matrix during electrophoresis?

    <p>Because they have a smaller mass-to-charge ratio</p> Signup and view all the answers

    What is the purpose of using a standard molecular weight marker in gel electrophoresis?

    <p>To estimate the size in bases or base pairs of the test nucleic acids</p> Signup and view all the answers

    Which of the following is a characteristic of polyacrylamide gels?

    <p>More precisely designed for high-resolution separation</p> Signup and view all the answers

    During electrophoresis, which pole do negatively charged nucleic acids move towards?

    <p>The anode</p> Signup and view all the answers

    What determines the proper type and concentration of gel to be used in electrophoresis?

    <p>The expected sizes of the nucleic acids to be separated</p> Signup and view all the answers

    Why are agarose gels less expensive and less toxic to use than polyacrylamide gels?

    <p>Because they are natural polymers</p> Signup and view all the answers

    What is the primary function of oligonucleotide primers in a PCR reaction?

    <p>To ensure the specificity of the PCR reaction</p> Signup and view all the answers

    What is the typical temperature range for the annealing step in a standard PCR cycle?

    <p>50°C–70°C</p> Signup and view all the answers

    What is the purpose of the initial 5- to 15-minute incubation at the denaturation temperature in some PCR protocols?

    <p>To activate specialized DNA polymerases</p> Signup and view all the answers

    What is the typical length of PCR products?

    <p>50 to more than 1,000 bp</p> Signup and view all the answers

    What is the purpose of the final 7- to 10-minute step at the extension temperature in some PCR protocols?

    <p>To ensure complete copying of the products</p> Signup and view all the answers

    What is the effect of ramp speed on PCR amplification?

    <p>It increases the efficiency of the amplification</p> Signup and view all the answers

    What is the purpose of the denaturation step in a standard PCR cycle?

    <p>To separate the double-stranded DNA into single strands</p> Signup and view all the answers

    What is the instrument used to carry out the amplification program in PCR?

    <p>Thermal cycler</p> Signup and view all the answers

    What is the typical number of cycles in a PCR amplification program?

    <p>20 to 50 cycles</p> Signup and view all the answers

    At what temperature is the extension step typically carried out in a standard PCR cycle?

    <p>68°C–72°C</p> Signup and view all the answers

    What is the primary purpose of the internal amplification control in qPCR and RT-qPCR?

    <p>To verify that negative results are true negatives and not due to amplification failure</p> Signup and view all the answers

    What is the primary difference between qPCR and digital droplet PCR?

    <p>The method of quantification of the target DNA</p> Signup and view all the answers

    What is the purpose of the limiting dilution of sample template molecules in digital droplet PCR?

    <p>To prepare individual droplets of reaction buffer in oil</p> Signup and view all the answers

    What is the primary application of qPCR in the detection of microorganisms?

    <p>To detect microorganisms that are difficult or dangerous to culture in the laboratory</p> Signup and view all the answers

    What is the function of the fluorescent dye in the capillary gel electrophoresis detection of PCR products?

    <p>To label the PCR primer</p> Signup and view all the answers

    What is the advantage of multiplex qPCR methods?

    <p>To assess multiple targets simultaneously</p> Signup and view all the answers

    What is the primary difference between the lag phase and the log phase in the PCR curve?

    <p>The rate of PCR product accumulation</p> Signup and view all the answers

    What is the purpose of the standard curve in qPCR?

    <p>To convert the Ct value to the number of DNA copies in the starting sample</p> Signup and view all the answers

    What is the primary advantage of digital droplet PCR over qPCR?

    <p>It provides absolute quantification</p> Signup and view all the answers

    What is the function of the emulsion formed in digital droplet PCR?

    <p>To act as individual reaction chambers</p> Signup and view all the answers

    What is the primary purpose of gel or capillary gel electrophoresis in a PCR procedure?

    <p>To visualize the products of the PCR reaction</p> Signup and view all the answers

    What is the role of reverse transcriptase in RT-PCR?

    <p>To synthesize cDNA from RNA</p> Signup and view all the answers

    What is the purpose of the 3′ end of the primer in SSP-PCR?

    <p>To anneal to the template on a potentially mutated or polymorphic base pair</p> Signup and view all the answers

    What is the advantage of using probes in qPCR compared to SYBR green?

    <p>Probes provide higher specificity for the intended product</p> Signup and view all the answers

    What is the purpose of the 5′ end of the primer in PCR?

    <p>To allow for the attachment of noncomplementary sequences containing restriction enzyme recognition sites</p> Signup and view all the answers

    What is the difference between RT-PCR and qPCR?

    <p>RT-PCR is used for qualitative detection, while qPCR is used for quantitative detection</p> Signup and view all the answers

    What is the purpose of FRET, TaqMan, molecular beacon, and scorpion probes in qPCR?

    <p>To hybridize to the template by sequence complementarity and generate fluorescent signals</p> Signup and view all the answers

    What is the advantage of using qPCR compared to standard PCR?

    <p>qPCR allows for the quantification of starting material</p> Signup and view all the answers

    What is the purpose of SYBR green in qPCR?

    <p>To detect the accumulation of PCR product in real time</p> Signup and view all the answers

    What is the purpose of sequence-specific primer PCR (SSP-PCR)?

    <p>To detect mutations and polymorphisms</p> Signup and view all the answers

    What is the primary function of RNase II in the process of probe amplification?

    <p>To degrade hybridized RNA, leaving single-stranded DNA</p> Signup and view all the answers

    In the process of strand displacement amplification, what is the function of the outer primer?

    <p>To bind to the target DNA and extend, displacing the product formed by the simultaneous extension of the inner primer</p> Signup and view all the answers

    What is the primary advantage of loop-mediated isothermal amplification (LAMP)?

    <p>Ability to amplify target DNA in a short amount of time</p> Signup and view all the answers

    In molecular inversion probe (MIP) amplification, what happens to the probe ends when they bind to target sequences?

    <p>They are brought together and ligated to form circles</p> Signup and view all the answers

    What is the primary function of branched DNA (bDNA) amplification?

    <p>To capture the target nucleic acid and bind to multiple reporter molecules</p> Signup and view all the answers

    What is the main advantage of branched DNA (bDNA) amplification over methods using a single probe or primer?

    <p>Higher specificity due to the use of multiple probes</p> Signup and view all the answers

    What is the primary function of DNA?

    <p>To store genetic information</p> Signup and view all the answers

    What is the primary difference between probe amplification and signal amplification?

    <p>Probe amplification amplifies the target DNA sequence, while signal amplification amplifies the signal bound to the target</p> Signup and view all the answers

    What is the purpose of the restriction enzyme in strand displacement amplification (SDA)?

    <p>To form a nick (cutting only one strand of the double helix) in the double-stranded DNA segment</p> Signup and view all the answers

    Which of the following sequencing methods uses modified nucleotide bases called dideoxynucleotide triphosphates (ddNTPs)?

    <p>Chain termination sequencing</p> Signup and view all the answers

    What is the primary application of molecular inversion probe (MIP) amplification?

    <p>All of the above</p> Signup and view all the answers

    What is the result of the sequencing reaction in Sanger sequencing?

    <p>A collection of fragments of various sizes</p> Signup and view all the answers

    What is the advantage of using multiple probes in branched DNA (bDNA) amplification?

    <p>Enhanced specificity due to the use of multiple probes</p> Signup and view all the answers

    What is the purpose of using fluorescent labels in Sanger sequencing?

    <p>To distinguish between the different ddNTPs</p> Signup and view all the answers

    What is the purpose of resolving the DNA ladder by gel or capillary gel electrophoresis?

    <p>To separate the DNA fragments based on their size</p> Signup and view all the answers

    What is the term for the collection of fluorescent peaks obtained by capillary gel electrophoresis?

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

    What is the purpose of the primer pair in Sanger sequencing?

    <p>To outline the target DNA</p> Signup and view all the answers

    What is the reason why synthesis will stop if a ddNTP is incorporated into the growing DNA chain?

    <p>Because the ddNTP lacks an OH group at the 3′ carbon of the deoxyribose sugar</p> Signup and view all the answers

    What is the advantage of capillary electrophoresis over gel electrophoresis?

    <p>It is faster and more automated</p> Signup and view all the answers

    What is the purpose of bDNA signal amplification assay?

    <p>To detect and quantify hepatitis B virus, HCV, and HIV-1</p> Signup and view all the answers

    What is the primary purpose of including positive and negative controls in in situ hybridization (ISH) testing?

    <p>To ensure the accuracy of the results and verify tissue preparation</p> Signup and view all the answers

    What is the main function of programmed cell death ligand (PD-L1) in the immune response?

    <p>To suppress the adaptive immune response by binding to programmed cell death protein 1 (PD-1) on activated lymphocytes and dendritic cells</p> Signup and view all the answers

    What is the primary difference between fluorescence in situ hybridization (FISH) and in situ hybridization (ISH) methods?

    <p>FISH requires specialized microscopes equipped to detect fluorescent signals, whereas ISH does not</p> Signup and view all the answers

    What is the purpose of using reference probes in fluorescence in situ hybridization (FISH) methods?

    <p>To identify the chromosomes of interest while assessing deletion or amplification</p> Signup and view all the answers

    What is the term used to describe the sensitivity of in situ hybridization (ISH) methods to the buffer and temperature conditions of hybridization?

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

    What is the main advantage of using array methods in molecular diagnostics?

    <p>They can detect multiple or complex genetic abnormalities</p> Signup and view all the answers

    What is the primary purpose of amplification methods in molecular diagnostics?

    <p>To copy nucleic acids for further analysis</p> Signup and view all the answers

    Who is credited with the development of the in vitro PCR, which greatly facilitated and broadened the potential applications of gene amplification?

    <p>Kerry Mullis</p> Signup and view all the answers

    What is the primary purpose of using immunohistochemistry in in situ hybridization (ISH) testing?

    <p>To detect the presence of clinically significant protein targets, such as those expressed by tumor cells</p> Signup and view all the answers

    What is the term used to describe the process of detecting targets in place as they appear in tissues, cells, and subcellular structures?

    <p>In situ hybridization (ISH)</p> Signup and view all the answers

    What is the primary mechanism by which TaqMan probes generate a signal?

    <p>The target DNA is copied, releasing the reporter dye from the quencher.</p> Signup and view all the answers

    What is the key difference between molecular beacons and TaqMan probes?

    <p>Molecular beacons have a quencher and a reporter dye, but they are not separated in the presence of target sequences.</p> Signup and view all the answers

    What is the purpose of the scorpion probe?

    <p>To generate a signal as the target DNA is copied, with the reporter covalently attached to the product.</p> Signup and view all the answers

    What is the primary advantage of digital droplet PCR over qPCR?

    <p>It is more sensitive and can detect smaller changes in target number.</p> Signup and view all the answers

    What is the primary mechanism of transcription-based amplification systems?

    <p>A complementary DNA copy is synthesized from the target RNA, and then transcription of the cDNA produces millions of copies of RNA products.</p> Signup and view all the answers

    What is the primary advantage of targeting RNA in transcription-based amplification systems?

    <p>It is more sensitive than targeting DNA because each microorganism makes multiple copies of RNA.</p> Signup and view all the answers

    What is the primary application of transcription-mediated amplification (TMA)?

    <p>Detection of RNA viruses, such as HCV and HIV.</p> Signup and view all the answers

    What is the purpose of the cDNA:RNA hybrid in transcription-mediated amplification?

    <p>To form a template for the synthesis of more cDNA.</p> Signup and view all the answers

    What is the advantage of TMA over PCR?

    <p>TMA is an isothermal process, which does not involve the repeated heating and cooling required for PCR.</p> Signup and view all the answers

    What is the primary application of digital PCR in infectious disease?

    <p>Detection of rare mutations in infectious diseases.</p> Signup and view all the answers

    What is the main purpose of Sanger sequencing in genetics and oncology?

    <p>To identify gene abnormalities in inherited diseases</p> Signup and view all the answers

    Why is pyrosequencing less sensitive than other methods for detecting DNA sequence changes?

    <p>Because it is affected by the heterozygous form of the alteration</p> Signup and view all the answers

    What is the purpose of sequencing the complementary strand of the DNA in Sanger sequencing?

    <p>To confirm the alteration detected in the original sequence</p> Signup and view all the answers

    What is the advantage of pyrosequencing over Sanger sequencing?

    <p>It is less labor-intensive and more convenient for short sequence analyses</p> Signup and view all the answers

    What is the purpose of the sequencing primer in pyrosequencing?

    <p>To initiate the DNA synthesis reaction</p> Signup and view all the answers

    What is the result of the pyrosequencing reaction?

    <p>A pyrogram of luminescent peaks</p> Signup and view all the answers

    Why is pyrosequencing used in genetics and oncology?

    <p>To sequence targeted areas of known gene locations</p> Signup and view all the answers

    What is the limitation of pyrosequencing compared to Sanger sequencing?

    <p>It produces shorter sequence reads</p> Signup and view all the answers

    What is the purpose of adenosine 5′ phosphosulfate (APS) in pyrosequencing?

    <p>To convert pyrophosphate to ATP</p> Signup and view all the answers

    What is the advantage of using some instruments developed for genomic or next-generation sequencing (NGS) in pyrosequencing?

    <p>They utilize the pyrosequencing chemistry</p> Signup and view all the answers

    What was the primary goal of developing Next-Generation Sequencing (NGS) technologies?

    <p>To sequence the human genome for a minimal cost of less than $1,000</p> Signup and view all the answers

    What is a characteristic of NGS procedures?

    <p>Starting with short DNA templates, usually less than 500 bp</p> Signup and view all the answers

    Which of the following NGS technologies has been most frequently applied to clinical applications?

    <p>Pyrosequencing, reversible dye sequencing, and ion conductance sequencing</p> Signup and view all the answers

    What was the primary challenge that was not initially included in the sequencing cost of NGS technologies?

    <p>The cost of interpreting the data</p> Signup and view all the answers

    What is the primary difference between NGS and traditional Sanger sequencing?

    <p>The number of genomes that can be sequenced simultaneously</p> Signup and view all the answers

    What is the term for the goal of sequencing the human genome for a minimal cost of less than $1,000?

    <p>The $1,000 genome</p> Signup and view all the answers

    What is the primary benefit of using large databases of somatic variant data in cancer diagnosis and treatment?

    <p>To refine interpretation of newly collected data</p> Signup and view all the answers

    What is the primary advantage of NGS technologies over traditional Sanger sequencing?

    <p>Lower cost of sequencing</p> Signup and view all the answers

    What is the name of the project that has sequenced thousands of genomes using NGS technologies?

    <p>The 1,000 Genome Project</p> Signup and view all the answers

    What is the primary advantage of using NGS over Sanger sequencing?

    <p>NGS provides higher coverage of regions of interest</p> Signup and view all the answers

    What is the purpose of using quality, variant type, and allele frequency parameters in filtering?

    <p>To identify the most biologically significant variants</p> Signup and view all the answers

    Why is adequate coverage of regions of interest required in NGS?

    <p>To provide sufficient data for confident base calling</p> Signup and view all the answers

    What is the primary purpose of annotating variants using historical sequence database information?

    <p>To determine the medical significance of variants</p> Signup and view all the answers

    What is the primary benefit of using a standard nomenclature system for sequence information?

    <p>To facilitate clear communication and organized storage of sequence data</p> Signup and view all the answers

    What is the primary purpose of using targeted gene panels in NGS?

    <p>To produce a finished report with variants identified automatically</p> Signup and view all the answers

    Why are errors more common in NGS compared to Sanger sequencing?

    <p>Library preparation is more error-prone in NGS</p> Signup and view all the answers

    What is the primary purpose of using FASTQ files in NGS?

    <p>To include a quality symbol for each base</p> Signup and view all the answers

    What is the primary benefit of using bioinformatics in molecular analyses?

    <p>To facilitate the interpretation of large amounts of data</p> Signup and view all the answers

    What is the primary purpose of comparing the sequence of a DNA sample to a reference sequence in NGS?

    <p>To identify variants or polymorphisms in the sample</p> Signup and view all the answers

    What is the minimum coverage required to call rare variants in NGS?

    <p>500X</p> Signup and view all the answers

    What is the primary application of targeted NGS in the clinical laboratory?

    <p>HLA typing, oncology, and genetics</p> Signup and view all the answers

    What is the purpose of the Cancer Genome Atlas (TCGA) program?

    <p>To understand the genetic basis of cancer and improve diagnosis and treatment</p> Signup and view all the answers

    What is the term for the number of mutations per megabase of sequenced DNA in a tumor?

    <p>Tumor mutational burden</p> Signup and view all the answers

    What is the primary advantage of NGS over other sequencing technologies?

    <p>Ability to investigate all known genetic loci</p> Signup and view all the answers

    What is the purpose of sequencing gene panels in NGS?

    <p>To identify genetic variants in a specific set of genes</p> Signup and view all the answers

    What is the term for the percentage of sequences carrying a variant in NGS?

    <p>Allele frequency</p> Signup and view all the answers

    What is the primary advantage of NGS in HLA typing?

    <p>Ability to resolve ambiguities</p> Signup and view all the answers

    What is the primary purpose of using PCR primers or probes in NGS?

    <p>To select and copy targeted regions for sequencing</p> Signup and view all the answers

    What is the primary factor that limits the degree of coverage used in NGS?

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

    What is the function of the bar code or index in NGS?

    <p>To identify each sample and gene region in the library</p> Signup and view all the answers

    What is the purpose of emulsion PCR in NGS?

    <p>To generate a library of PCR products attached to beads</p> Signup and view all the answers

    What is the mechanism of detection in ion conductance sequencing?

    <p>Release of hydrogen ions and detection of pH change</p> Signup and view all the answers

    What is the purpose of bridge PCR in reversible dye terminator technology?

    <p>To form clusters of templates across the flow cell</p> Signup and view all the answers

    What is the advantage of ion conductance sequencing over other methods?

    <p>It is faster and does not require optical sensing</p> Signup and view all the answers

    What is the purpose of adapters in NGS?

    <p>To carry PCR primer-binding sites and allow unbiased amplification</p> Signup and view all the answers

    What is the purpose of the sequencing reagents in pyrosequencing?

    <p>To release PPi and generate light signals</p> Signup and view all the answers

    What is the advantage of using multiple samples in a single run of NGS?

    <p>It decreases the cost and time required for sequencing</p> Signup and view all the answers

    What is the purpose of the picoplate in pyrosequencing?

    <p>To carry out independent sequencing reactions in hundreds of thousands of wells</p> Signup and view all the answers

    What is the primary function of flow cytometry in the clinical laboratory?

    <p>To monitor HIV infection status and enumerate hematopoietic stem cells</p> Signup and view all the answers

    What is the primary purpose of using fluorescent-labeled antibodies or probes in flow cytometry?

    <p>To identify specific surface or cytoplasmic proteins</p> Signup and view all the answers

    What is a characteristic of fluorochromes used in flow cytometry?

    <p>They absorb light across a spectrum of wavelengths and emit light of lower energy across a spectrum of longer wavelengths</p> Signup and view all the answers

    What is the primary application of flow cytometry in the diagnosis of diseases?

    <p>In the diagnosis of leukemia and lymphoma</p> Signup and view all the answers

    What is the primary advantage of using multiple fluorochromes in flow cytometry?

    <p>It allows for the analysis of multiple cell properties simultaneously</p> Signup and view all the answers

    What is the primary function of laser light in flow cytometry?

    <p>To excite fluorochromes and detect cell properties</p> Signup and view all the answers

    What is the primary application of flow cytometry in transplantation?

    <p>To detect human leukocyte antigen (HLA) antibodies</p> Signup and view all the answers

    What is the primary advantage of using flow cytometry in cell analysis?

    <p>It allows for the analysis of a large number of cells simultaneously</p> Signup and view all the answers

    What is the primary purpose of using optical filters in flow cytometry?

    <p>To maximize collection of light from specific fluorochromes</p> Signup and view all the answers

    What is the effect of using Ficoll-Hypaque density gradient centrifugation on some cell populations?

    <p>Selective loss of some cell populations</p> Signup and view all the answers

    What is the purpose of using erythrocyte lysis techniques in sample preparation for flow cytometry?

    <p>To destroy erythrocytes while leaving WBCs intact</p> Signup and view all the answers

    What is the typical range of digital signals measured in flow cytometry?

    <p>1 to 256 channels</p> Signup and view all the answers

    What is the purpose of using a marker in flow cytometry data analysis?

    <p>To differentiate cells with low levels of fluorescence from cells with high levels of fluorescence</p> Signup and view all the answers

    What is the primary purpose of using fiber-optic cables in flow cytometry?

    <p>To direct light to the photomultiplier tubes</p> Signup and view all the answers

    What is the effect of using heparin as an anticoagulant in sample preparation for flow cytometry?

    <p>Improved stability of samples for up to 48 hours</p> Signup and view all the answers

    What is the primary purpose of using EDTA as an anticoagulant in sample preparation for flow cytometry?

    <p>To collect whole blood samples for analysis within 30 hours</p> Signup and view all the answers

    What is the typical number of events collected for each sample in flow cytometry data acquisition?

    <p>10,000 to 20,000 events</p> Signup and view all the answers

    What is the primary purpose of using a single-parameter histogram in flow cytometry data analysis?

    <p>To plot a chosen parameter versus the number of events</p> Signup and view all the answers

    What is the primary advantage of flow cytometry?

    <p>It enables the accurate detection of cells that are present in very small numbers.</p> Signup and view all the answers

    What is the purpose of the fluidics system in a flow cytometer?

    <p>To ensure that cells pass through the laser light one cell at a time.</p> Signup and view all the answers

    What determines which fluorochromes can be used in an assay?

    <p>The wavelength of the laser light.</p> Signup and view all the answers

    What is the primary defect in patients with paroxysmal nocturnal hemoglobinuria (PNH)?

    <p>Lack of GPI anchor proteins on the cell surface</p> Signup and view all the answers

    Which of the following tests can detect minimal residual disease (MRD) in patients with leukemia or lymphoma?

    <p>Flow cytometry</p> Signup and view all the answers

    What is the difference between forward scatter and side scatter?

    <p>Forward scatter measures size, while side scatter measures granularity.</p> Signup and view all the answers

    What is the purpose of using fluorescent-labeled antibodies in flow cytometry?

    <p>To detect the presence of specific antigens on the surface of cells.</p> Signup and view all the answers

    What is the primary application of flow cytometry in transplant immunology?

    <p>HLA typing and cross-matching</p> Signup and view all the answers

    What is the advantage of using multiple lasers in a flow cytometer?

    <p>It enables the analysis of up to five fluorochromes at a time.</p> Signup and view all the answers

    What is the advantage of using monoclonal antibodies over polyclonal antibodies in immunophenotyping?

    <p>Monoclonal antibodies are more specific</p> Signup and view all the answers

    What is the primary function of cytometric bead arrays?

    <p>Identifying multiple analytes simultaneously</p> Signup and view all the answers

    What is the purpose of the optics and photodetectors in a flow cytometer?

    <p>To detect the light scatter and fluorescence signals generated by the cells.</p> Signup and view all the answers

    What is the term for the parameters that are intrinsic to the cell?

    <p>Intrinsic parameters.</p> Signup and view all the answers

    Which of the following is an application of flow cytometry in molecular diagnostics?

    <p>Detecting viral nucleic acids</p> Signup and view all the answers

    What is the advantage of using multicolor analysis in flow cytometry?

    <p>It enables the analysis of multiple extrinsic parameters simultaneously.</p> Signup and view all the answers

    What is the primary advantage of flow cytometry over traditional serological methods?

    <p>Faster and more accurate results</p> Signup and view all the answers

    What is the purpose of analyzing the data generated by the flow cytometer in conjunction with forward scatter and side scatter?

    <p>To enable more accurate subtyping of cells.</p> Signup and view all the answers

    What is the purpose of fluorescent labels in bead array assays?

    <p>To detect analytes</p> Signup and view all the answers

    Which of the following is a clinical application of flow cytometry?

    <p>All of the above</p> Signup and view all the answers

    What is the primary purpose of a gate in a dual-parameter dot plot?

    <p>To filter out debris and isolate subpopulations of cells of interest</p> Signup and view all the answers

    What is the primary advantage of using flow cytometry in DNA analysis?

    <p>All of the above</p> Signup and view all the answers

    What is the advantage of using a single-platform analysis over a dual-platform analysis?

    <p>It eliminates the potential for added error associated with the use of two distinct methods</p> Signup and view all the answers

    What is the primary purpose of immunophenotyping in the clinical laboratory?

    <p>To characterize and enumerate normal lymphocytes</p> Signup and view all the answers

    What is the primary difference between quadrants 1 and 2 in a dual-parameter dot plot?

    <p>Quadrant 1 consists of cells that are positive for fluorescence on the y-axis and negative for fluorescence on the x-axis, while quadrant 2 consists of cells that are positive for fluorescence on both the x- and y-axes</p> Signup and view all the answers

    What is the primary purpose of the computer and specialized software in a dual-parameter dot plot analysis?

    <p>To calculate the percentage of cells in each quadrant</p> Signup and view all the answers

    What is the primary advantage of using bead-based methods over volumetric methods in single-platform analysis?

    <p>Bead-based methods provide a more accurate absolute count of a particular cell type</p> Signup and view all the answers

    What is the primary purpose of detailed phenotypic analysis in the clinical laboratory?

    <p>To determine the degree of differentiation and activation of a specific cell population</p> Signup and view all the answers

    What is the primary difference between nonmalignant immunophenotyping and malignant immunophenotyping?

    <p>Nonmalignant immunophenotyping is used to characterize and enumerate normal lymphocytes, while malignant immunophenotyping is used to diagnose malignant disorders</p> Signup and view all the answers

    What is the primary purpose of using a known quantity of fluorescent beads in a bead-based method?

    <p>To provide a simple mathematic calculation to directly determine the absolute WBC numbers</p> Signup and view all the answers

    What is the primary advantage of using a single-platform analysis over a traditional method?

    <p>It eliminates the potential for added error associated with the use of two distinct methods</p> Signup and view all the answers

    What is the primary purpose of immunophenotyping by flow cytometry in leukemia and lymphoma management?

    <p>To identify the lineage of hematopoietic malignancies</p> Signup and view all the answers

    Which of the following markers is typically paired with CD20 in the analysis of chronic lymphocytic leukemia (CLL)?

    <p>CD5</p> Signup and view all the answers

    What is the significance of CD45 expression levels in flow cytometry analysis?

    <p>It is used to differentiate various populations of WBCs</p> Signup and view all the answers

    What is the primary application of flow cytometry in the evaluation of HIV disease?

    <p>To classify stages of HIV disease and determine treatment options</p> Signup and view all the answers

    What is the characteristic of neutrophils from patients with chronic granulomatous disease (CGD)?

    <p>They are unable to oxidize the dye dihydrorhodamine 123</p> Signup and view all the answers

    What is the purpose of examining the FSC and SSC plot in flow cytometry analysis?

    <p>To ensure that a population is not being missed</p> Signup and view all the answers

    What is the significance of CD4+ T-cell numbers in HIV-infected patients?

    <p>They are used to classify stages of HIV disease and determine treatment options</p> Signup and view all the answers

    What is the primary purpose of immunophenotyping in the evaluation of immunodeficiency?

    <p>To compare patient values with reference ranges</p> Signup and view all the answers

    What is the significance of CD8+ T-cell levels in HIV-infected patients?

    <p>They are expanded during the early course of HIV disease</p> Signup and view all the answers

    What is the characteristic of blasts in terms of CD45 expression?

    <p>They have low levels of CD45 expression</p> Signup and view all the answers

    What was the primary motivation for the development of immunoassay analyzers?

    <p>To create an automated system capable of reducing or eliminating manual tasks</p> Signup and view all the answers

    What is a benefit of immunoassay automation in addition to reducing error and increasing accuracy?

    <p>The ability to provide more services with less staff</p> Signup and view all the answers

    What is a challenge of selecting an immunoassay analyzer for a laboratory?

    <p>The difficulty of determining which analyzer is best suited for a laboratory's needs</p> Signup and view all the answers

    What is a benefit of immunoassay automation in terms of reagent management?

    <p>The longer shelf life of reagents</p> Signup and view all the answers

    What is a common type of analyte that can be measured using immunoassay automation?

    <p>Low-level peptides such as peptide hormones</p> Signup and view all the answers

    How many different types of automated immunoassay analyzers are available?

    <p>More than 60</p> Signup and view all the answers

    What is a consequence of eliminating manual steps in immunoassay testing?

    <p>The decrease in the likelihood of error</p> Signup and view all the answers

    What is a potential benefit of immunoassay automation in terms of sample management?

    <p>Sample delivery with bar codes for better sample identification</p> Signup and view all the answers

    What is a primary limitation of batch analyzers?

    <p>They can only perform one type of analysis at a time</p> Signup and view all the answers

    What is the purpose of clot detectors in stainless-steel probes?

    <p>To reject samples with clots</p> Signup and view all the answers

    What is a common issue associated with reusable pipette probes?

    <p>Carryover or contamination of samples</p> Signup and view all the answers

    What is the primary purpose of reagent storage compartments in some analyzers?

    <p>To store reagents at a controlled temperature</p> Signup and view all the answers

    What is the purpose of magnetic stirring in analyzers?

    <p>To mix samples with reagents</p> Signup and view all the answers

    What is the primary purpose of timed incubation in analyzers?

    <p>To incubate samples at a controlled temperature</p> Signup and view all the answers

    What is the primary advantage of using batch analyzers in clinical laboratories?