Test Your PCR Expertise

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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?

About 109 copies of the target segment Signup and view all the answers

What is the estimated efficiency of amplification in PCR?

60% to 85% Signup and view all the answers

What is the purpose of a thermal cycler in PCR?

To shift the temperature up and down during different stages of the cycle Signup and view all the answers

What are some practical applications of PCR?

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 Signup and view all the answers

How long does each cycle of PCR take?

4 to 6 minutes Signup and view all the answers

What is the role of gel electrophoresis in PCR?

To characterize the amplified DNA by restriction endonuclease digestion Signup and view all the answers

What is the name of the inventor of PCR?

Kary B. Mullis Signup and view all the answers

What is the Sanger method used for?

Sequencing of DNA Signup and view all the answers

What are dideoxynucleoside triphosphates?

Analogues of deoxynucleoside triphosphates Signup and view all the answers

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

To label the dideoxynucleotides Signup and view all the answers

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

To determine the region of DNA to be sequenced Signup and view all the answers

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

It requires less DNA Signup and view all the answers

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

To amplify the reaction products Signup and view all the answers

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

It diminishes the chance of an error Signup and view all the answers

What is the difference between deoxynucleoside triphosphates and dideoxynucleoside triphosphates?

Dideoxynucleoside triphosphates have a hydroxyl group attached to C-3′ Signup and view all the answers

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

To separate the DNA strands Signup and view all the answers

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

Frederick Sanger Signup and view all the answers