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 (C)</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 (A)</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. (D)</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. (A)</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. (A)</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. (D)</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. (B)</p> Signup and view all the answers

Flashcards

Southern blotting purpose

Determine the presence of specific DNA sequences in samples.

Probe function

Identify DNA fragments that complement a specific probe.

DNA amount for digestion

Depends on the type of DNA sample being used.

DNA fragmentation purpose

To create DNA fragments for analysis.

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Edwin M. Southern

Inventor of Southern analysis in DNA technology.

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Purine stability

Purine bases are less likely to depurinate than pyrimidines.

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Capillary action in DNA transfer

Allows quick transfer of DNA fragments onto a membrane.

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0.4 M NaOH role

Enhances DNA binding to nylon membranes.

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Alkaline buffer for transfer

Used to increase binding of DNA onto membranes.

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20X SSC use

Improves stability and binding of DNA to nitrocellulose.

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