STBP2023 DNA Recombinant Technology I Southern Blotting
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Questions and Answers

What is the primary purpose of transferring DNA onto a membrane in Southern blotting?

  • To isolate the DNA from the sample
  • To determine the presence of a specific DNA sequence within a large, complex DNA sample (correct)
  • To amplify the DNA for further analysis
  • To visualize the DNA fragments under a microscope
  • What is the role of probe in Southern blotting?

  • To visualize the labeled DNA on X-ray film
  • To fragment the DNA using restriction enzymes
  • To identify which DNA fragments obtained from a digest of a larger DNA clone/genome that contains sequence complementary to a specific probe (correct)
  • To extract genomic DNA from the sample
  • What determines the amount of DNA required for restriction enzyme digestion in Southern blotting?

  • Type of DNA sample (e.g., plasmid DNA, genomic DNA) (correct)
  • The temperature at which the membrane is incubated
  • The concentration of the radioactive DNA used
  • The size of the membrane used
  • What is the purpose of DNA fragmentation using restriction endonucleases in Southern blotting?

    <p>To obtain DNA fragments for further analysis</p> Signup and view all the answers

    Who invented Southern analysis, a key technique in DNA recombinant technology?

    <p>Edwin M. Southern, an English Molecular Biologist</p> Signup and view all the answers

    What is the purpose of using purine bases instead of pyrimidines during DNA treatment with diluted acid?

    <p>Purine bases are less likely to be depurinated compared to pyrimidines.</p> Signup and view all the answers

    Why is capillary action used for the transfer of DNA onto the membrane?

    <p>Capillary action allows for the rapid transfer of DNA fragments.</p> Signup and view all the answers

    What is the role of 0.4 M NaOH in the transfer of DNA onto a nylon membrane?

    <p>It enhances the binding of DNA onto the nylon membrane.</p> Signup and view all the answers

    Why is an alkaline buffer used for transferring DNA onto a nylon membrane?

    <p>To enhance the binding of DNA onto the nylon membrane.</p> Signup and view all the answers

    What is the main reason for using 20X SSC for transferring DNA onto a nitrocellulose membrane?

    <p>To improve the stability and binding of DNA onto the nitrocellulose membrane.</p> Signup and view all the answers

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