Test Your PCR Expertise

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Questions and Answers

Which enzyme was used in the polymerase chain reaction (PCR) to extend the primer?

  • Helicase
  • RNA polymerase
  • DNA polymerase I (correct)
  • Ligase

What is the role of the thermostable DNA polymerase in PCR?

  • To anneal to the denatured single strands
  • To avoid repeated DNA polymerase I additions at each cycle (correct)
  • To extend the primer with RNA polymerase
  • To amplify the target sequence

What is the purpose of the oligonucleotide primer in PCR?

  • To sequence the amplified DNA
  • To anneal to each of the denatured single strands (correct)
  • To denature the linear duplex
  • To amplify the target sequence

What is the expected amplification yield after 30 cycles of PCR?

<p>About 109 copies of the target segment (D)</p> Signup and view all the answers

What is the estimated efficiency of amplification in PCR?

<p>60% to 85% (B)</p> Signup and view all the answers

What is the purpose of a thermal cycler in PCR?

<p>To shift the temperature up and down during different stages of the cycle (A)</p> Signup and view all the answers

What are some practical applications of PCR?

<p>Amplification of a segment of a large DNA molecule, rapid detection of pathogenic bacteria and viruses, detection of inborn errors of metabolism, detection of tumors (B)</p> Signup and view all the answers

How long does each cycle of PCR take?

<p>4 to 6 minutes (A)</p> Signup and view all the answers

What is the role of gel electrophoresis in PCR?

<p>To characterize the amplified DNA by restriction endonuclease digestion (A)</p> Signup and view all the answers

What is the name of the inventor of PCR?

<p>Kary B. Mullis (A)</p> Signup and view all the answers

What is the Sanger method used for?

<p>Sequencing of DNA (B)</p> Signup and view all the answers

What are dideoxynucleoside triphosphates?

<p>Analogues of deoxynucleoside triphosphates (C)</p> Signup and view all the answers

What is the purpose of the fluorescent dyes in the Sanger method?

<p>To label the dideoxynucleotides (D)</p> Signup and view all the answers

What is the role of the oligonucleotide primer in the Sanger method?

<p>To determine the region of DNA to be sequenced (B)</p> Signup and view all the answers

What is the major advantage of dideoxy-terminator cycle sequencing over the standard Sanger method?

<p>It requires less DNA (C)</p> Signup and view all the answers

What is the function of the thermal cycler in dideoxy-terminator cycle sequencing?

<p>To amplify the reaction products (C)</p> Signup and view all the answers

What is the advantage of sequencing both strands of a double-stranded DNA in dideoxy-terminator cycle sequencing?

<p>It diminishes the chance of an error (C)</p> Signup and view all the answers

What is the difference between deoxynucleoside triphosphates and dideoxynucleoside triphosphates?

<p>Dideoxynucleoside triphosphates have a hydroxyl group attached to C-3′ (A)</p> Signup and view all the answers

What is the purpose of denaturation in dideoxy-terminator cycle sequencing?

<p>To separate the DNA strands (C)</p> Signup and view all the answers

What is the name of the scientist who developed the Sanger method?

<p>Frederick Sanger (A)</p> Signup and view all the answers

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