Recombinant DNA Technology Process
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Questions and Answers

What is the first step in Recombinant DNA technology?

  • Isolation of Genetic Material (correct)
  • Insertion of Recombinant DNA Into Host
  • Amplifying the gene copies through PCR
  • Cutting the gene at the recognition sites

    • What is the role of restriction enzymes in Recombinant DNA technology?

  • Determining the gene insertion location into the vector genome (correct)
  • Amplifying the gene copies
  • Isolating the genetic material
  • Transforming host cells

    • What is the purpose of ligation in DNA cloning?

  • Joining a cut fragment of DNA with a vector using DNA ligase (correct)
  • Isolating Genetic Material
  • Inserting Recombinant DNA into host
  • Determining gene insertion location

    • What term describes the process of introducing recombinant DNA into a recipient host cell?

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

    What is the outcome of effectively transforming cells/organisms in Recombinant DNA technology?

    <p>Expression of manufactured protein</p> Signup and view all the answers

    What does the term 'clone' refer to in DNA cloning?

    <p>A group of individual cells descended from one progenitor</p> Signup and view all the answers

    What is the technology used for producing artificial DNA by combining genetic materials from different sources?

    <p>Genetic Engineering</p> Signup and view all the answers

    Who discovered restriction enzymes in the year 1968?

    <p>Werner Arber</p> Signup and view all the answers

    What is the role of ligases in recombinant DNA technology?

    <p>Help to bind DNA fragments</p> Signup and view all the answers

    How do restriction endonucleases cut DNA?

    <p>Within the DNA strand</p> Signup and view all the answers

    What is the purpose of DNA primers in the PCR process?

    <p>To promote the synthesis of a complementary DNA strand</p> Signup and view all the answers

    What is the main function of Exonucleases in recombinant DNA technology?

    <p>Remove nucleotides from the ends of DNA strands</p> Signup and view all the answers

    During which PCR machine step does denaturation of double-stranded DNA occur?

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

    What do sticky ends refer to in recombinant DNA technology?

    <p>Regions where DNA fragments can bind due to complementary base pairing</p> Signup and view all the answers

    What enzyme aids in the synthesis of a complementary strand of DNA in the PCR process?

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

    What happens to the original DNA sample during the denaturation step in the PCR machine?

    <p>The DNA is heated to at least 94°C</p> Signup and view all the answers

    Which step in the PCR process involves cooling the sample mixture to allow DNA primers and DNA polymerase to bind to individual strands of DNA?

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

    What is the function of the nucleotide solution mix in the PCR process?

    <p>To build duplicate DNA strands</p> Signup and view all the answers

    What is the term used to describe the process of forming a new complementary strand of DNA after the nucleotides pair with individual DNA strands?

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

    Which temperature range is involved in the cycling process of heating and cooling during DNA amplification?

    <p>95°C to 50 to 60°C</p> Signup and view all the answers

    How many times is the cycling process of heating and cooling repeated using the thermal cycler?

    <p>35 to 40 times</p> Signup and view all the answers

    What happens to the resulting DNA sequence each time the heating/cooling cycle is conducted by the cycler?

    <p>It is doubled</p> Signup and view all the answers

    Why can millions of copies of DNA be formed after 35 doubling cycles?

    <p>Due to extension of the DNA strands</p> Signup and view all the answers

    What makes the work of ligases easy while binding the desired gene to the vector?

    <p>Cutting the desired gene and vector with different restriction enzymes</p> Signup and view all the answers

    Which of the following is not a component of vectors used in recombinant DNA technology?

    <p>Restriction enzymes</p> Signup and view all the answers

    What feature of plasmids and bacteriophages makes them popular as vectors in recombinant DNA technology?

    <p>High copy number</p> Signup and view all the answers

    Which step in the process of recombinant DNA technology involves introducing the recombinant DNA into the host organism?

    <p>Using microinjection</p> Signup and view all the answers

    What is the main role of the host organism in recombinant DNA technology?

    <p>Carrying the desired gene forward</p> Signup and view all the answers

    What is the sequence of nucleotides within a vector from where replication starts known as?

    <p>Origin of replication</p> Signup and view all the answers

    Study Notes

    Recombinant DNA Technology

    • Also known as Genetic Engineering
    • Involves combining different genetic materials (DNA) from different sources to produce artificial DNA

    History of Recombinant DNA Technology

    • Emerged with the discovery of restriction enzymes in 1968 by Swiss microbiologist Werner Arber

    Tools of Recombinant DNA Technology

    • Enzymes: restriction enzymes, polymerases, and ligases
    • Restriction enzymes: cut the DNA at specific points, producing sticky ends
    • Polymerases: synthesize DNA
    • Ligases: bind DNA molecules
    • Vectors: carry and integrate the desired gene into the host organism
    • Plasmids and bacteriophages are commonly used vectors due to their high copy number

    Process of Recombinant DNA Technology

    • Step 1: Isolation of genetic material
    • Step 2: Cutting the gene at recognition sites using restriction enzymes
    • Step 3: Amplifying the gene copies through PCR
    • Step 4: Ligation of DNA molecules
    • Step 5: Insertion of recombinant DNA into the host

    PCR (Polymerase Chain Reaction)

    • A process to amplify a single copy of DNA into thousands to millions of copies
    • Requires four components: DNA or RNA sample, DNA primers, DNA polymerase, and nucleotide solution mix
    • PCR process has 4 steps: collection, preparation, amplification, and post-PCR clean-up
    • PCR machine steps: denaturation, annealing, and extension
    • Denaturation: heat breaks hydrogen bonds, separating DNA into single strands
    • Annealing: cooling allows DNA primers and polymerase to bind to individual strands
    • Extension: new complementary strand of DNA is formed, resulting in a new duplicate double-stranded DNA molecule

    DNA Cloning

    • A clone is a cluster of individual entities or cells that are descended from one progenitor
    • Recombinant DNA is introduced into a recipient host cell, which gets multiplied and expressed in the form of the manufactured protein under optimal conditions

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    Description

    Learn about the key steps involved in Recombinant DNA technology, including isolation of genetic material, cutting the gene at recognition sites using restriction enzymes, and amplifying gene copies through Polymerase Chain Reaction (PCR).

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