Gene amplification
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Questions and Answers

What is gene amplification?

  • A random change in the number of genes present in a cell
  • An increase in the copy of a particular gene without an increase in the number of copies of other genes (correct)
  • An increase in the number of genes present in a cell
  • A decrease in the copy of a particular gene without affecting the number of copies of other genes

    • How can gene amplification occur naturally?

  • Through gene deletion, replication and repair slippage, crossing-over between homologous chromosomes, aneuploidy, and polyploidy
  • Through gene translocation, replication and repair slippage, ectopic recombination, aneuploidy, and monoploidy
  • Through gene inversion, replication and repair slippage, crossing-over between non-homologous chromosomes, aneuploidy, and monoploidy
  • Through gene duplication, replication and repair slippage, ectopic recombination, aneuploidy, and polyploidy (correct)

    • What is the term for the piece of DNA or the gene that gets amplified naturally or artificially?

  • Exon
  • Codon
  • Transcript
  • Amplicon (correct)

    • How is gene amplification artificially done in a lab?

    <p>By polymerase chain reaction (PCR)</p> Signup and view all the answers

    What is the recommended range for GC content in primer sequences?

    <p>40-60%</p> Signup and view all the answers

    Why is it important to check primer specificity by BLAST?

    <p>To ensure the primers only bind to the target sequence</p> Signup and view all the answers

    What is the recommended length range for primer sequences?

    <p>18-25 bp</p> Signup and view all the answers

    What is the formula to calculate the melting temperature (Tm) of a primer?

    <p>Tm = (G + C) x 4 + (A + T) x 2</p> Signup and view all the answers

    What does the annealing temperature (Ta) depend directly on?

    <p>Length and GC composition of the primers</p> Signup and view all the answers

    What does the presence of a GC clamp in a primer sequence refer to?

    <p>Having G or C bases within the last five bases from the 5' end of the primer</p> Signup and view all the answers

    What is the recommended range for melting temperatures (Tm) of primer sequences?

    <p>50-60 °C</p> Signup and view all the answers

    Why is it important to know the product size of the amplified target sequence?

    <p>To ensure the target sequence is amplified and of the expected size</p> Signup and view all the answers

    Which technique utilizes RNA intermediates for gene amplification?

    <p>Transcription-mediated amplification (TMA)</p> Signup and view all the answers

    What is a specific advantage of Ligase chain reaction (LCR) over PCR?

    <p>Increased specificity for point mutations</p> Signup and view all the answers

    Which method is commonly used to produce billions of copies of a target DNA sequence in a short time?

    <p>Polymerase chain reaction (PCR)</p> Signup and view all the answers

    What governs the three distinct steps of denaturation, annealing, and extension in PCR?

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

    What determines the number of copies produced in PCR?

    <p>Number of cycles</p> Signup and view all the answers

    What is a key component required for PCR to target the DNA to be amplified?

    <p>Synthetic oligonucleotide primers</p> Signup and view all the answers

    What is the role of reverse transcriptase in Transcription-mediated amplification (TMA)?

    <p>Catalyzes RNA to DNA conversion</p> Signup and view all the answers

    What is the primary purpose of post-PCR analysis?

    <p>Primer design and PCR optimization</p> Signup and view all the answers

    What is the main advantage of Ligase chain reaction (LCR) over PCR?

    <p>Increased specificity for point mutations</p> Signup and view all the answers

    What is the primary function of Ligase chain reaction (LCR)?

    <p>Detection of point mutations</p> Signup and view all the answers

    Which enzyme is essential for the rapid amplification of target genes in Transcription-mediated amplification (TMA)?

    <p>RNA polymerase</p> Signup and view all the answers

    What is the main role of DNA polymerase in PCR?

    <p>Synthesizes new DNA strands</p> Signup and view all the answers

    What is the main purpose of gene amplification?

    <p>Disease diagnosis and understanding disease progression</p> Signup and view all the answers

    Which gene amplification technique is more specific than PCR and used to identify point mutations?

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

    What is the main role of transcription-mediated amplification (TMA)?

    <p>Rapidly amplifying target genes</p> Signup and view all the answers

    What is the main function of PCR in the lab?

    <p>Producing multiple copies of a desired gene</p> Signup and view all the answers

    What governs the three distinct steps of PCR: denaturation, annealing, and extension?

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

    How many copies of the target DNA can PCR produce in about 20 cycles?

    <p>About a million</p> Signup and view all the answers

    What is the main requirement for PCR to target the DNA to be amplified?

    <p>Specific DNA sequence information</p> Signup and view all the answers

    What is the mechanism of PCR primarily involved in?

    <p>Denaturation of template DNA, primer annealing, and DNA synthesis</p> Signup and view all the answers

    What does post-PCR analysis involve?

    <p>Primer design, primer specificity, PCR optimization, and visualization of results</p> Signup and view all the answers

    What determines the number of copies produced in PCR?

    <p>2n, where n is the number of cycles</p> Signup and view all the answers

    What is the main role of ligase chain reaction (LCR) in gene amplification?

    <p>Identifying point mutations</p> Signup and view all the answers

    What is the primary function of gene amplification in research and diagnosis?

    <p>To produce desired gene products</p> Signup and view all the answers

    Study Notes

    Gene Amplification Techniques: PCR, LCR, and TMA

    • Retrotransposition is a type of transposon that copies itself through RNA intermediates, and is commonly found in tumour cells.
    • Amplification of genes is useful for disease diagnosis, understanding disease progression, and can lead to drug resistance in cancer cells.
    • Artificial gene amplification is used in research, diagnosis, recombinant DNA technology, and to produce desired gene products.
    • Polymerase chain reaction (PCR) is a commonly used method to produce multiple copies of a desired gene in the lab, and is essential in modern science and biotechnology.
    • Ligase chain reaction (LCR) is another method of gene amplification, more specific than PCR, and widely used to ascertain point mutations such as in genetic diseases.
    • Transcription-mediated amplification (TMA) utilizes RNA polymerase and reverse transcriptase to rapidly amplify target genes.
    • PCR is a means to amplify a particular piece of DNA and can make billions of copies of a target sequence of DNA in a short time.
    • PCR requires specific DNA sequence information to target the DNA to be amplified and uses synthetic oligonucleotide primers.
    • PCR proceeds in three distinct steps governed by temperature: denaturation, annealing, and extension.
    • The mechanism of PCR involves denaturation of template DNA, primer annealing, and DNA synthesis by DNA polymerase.
    • PCR can produce about a million copies of the target in only 20 cycles, and the number of copies is determined by 2n, where n is the number of cycles.
    • Post-PCR analysis involves primer design, primer specificity, PCR optimization, and visualization of results using agarose gel electrophoresis.

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    Pr Lecture IVa_Gene Amplification and PCR_Lecture.pptx

    Description

    Explore gene amplification techniques like PCR, LCR, and TMA used in research, diagnosis, and biotechnology. Learn about the mechanisms, applications, and significance of amplifying genes in disease diagnosis and drug resistance.

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