chapter 9

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What is the main purpose of Polymerase Chain Reaction (PCR)?

To make billions of copies of a specific DNA sequence (correct)

To produce new DNA sequences

To identify different types of DNA

To analyze the structure of DNA molecules

What does 'in vitro' refer to in the context of PCR?

DNA modification in a controlled environment

DNA replication in a test tube (correct)

DNA replication in living cells

DNA sequencing in a laboratory setting

How does PCR relate to DNA Replication in cells?

It is a laboratory version of DNA Replication (correct)

It is an alternative process to DNA Replication

It is used to inhibit DNA Replication

It has no relation to DNA Replication

What is one of the applications of PCR mentioned in the text?

Detecting mutations in specific genes for genetic testing Signup and view all the answers

How does PCR contribute to forensic analysis?

Identifying and matching DNA samples from crime scenes to suspects Signup and view all the answers

In what way does PCR support drug development?

Identifying and validating drug targets and screening potential drug candidates Signup and view all the answers

What is the purpose of DNA Extraction Lysis in sample preparation?

To break cells and release DNA Signup and view all the answers

What is the role of Proteinase K in the DNA extraction process?

To dissolve cellular proteins Signup and view all the answers

What is the final step in DNA purification during sample preparation?

Eluting the DNA with a slightly alkaline buffer Signup and view all the answers

What does PCR copy from the sample?

A very specific sequence of genetic code from a template DNA Signup and view all the answers

What information is required for the amplification of a specific target sequence using PCR?

DNA sequence information which flanks the fragment of DNA to be amplified Signup and view all the answers

What is the role of PCR primers in the amplification process?

To target a specific sequence of genetic code for amplification Signup and view all the answers

What does PCR copy from the sample?

A very specific sequence of genetic code from a template DNA Signup and view all the answers

What information is required for the amplification of a specific target sequence using PCR?

DNA sequence information which flanks the fragment of DNA to be amplified Signup and view all the answers

What is the main purpose of Polymerase Chain Reaction (PCR)?

To amplify a specific target sequence of DNA Signup and view all the answers

What is the purpose of oligonucleotide primers in PCR?

To initiate DNA synthesis at a specific target sequence Signup and view all the answers

What is the role of DNA polymerase in PCR?

Adding nucleotides onto pre-existing 3'-OH groups to extend DNA strands Signup and view all the answers

Why are two oligonucleotide primers required in PCR amplification?

To allow amplification of both DNA strands Signup and view all the answers

Why are two primers required in a PCR reaction?

To amplify both strands of the double-stranded DNA Signup and view all the answers

What is the role of the forward primer in PCR?

It binds to the complementary reverse DNA strand Signup and view all the answers

What happens if only one primer is used in a PCR reaction?

Only one strand of the double-stranded DNA will be amplified Signup and view all the answers

What is the recommended GC content range for a good primer?

40-60% Signup and view all the answers

Why is it important for the 3’ of a primer to end in G or C?

To promote binding Signup and view all the answers

What impact do G and C bases have on the melting temperature (Tm) of primers?

They increase Tm Signup and view all the answers

At what temperature does denaturation of dsDNA usually occur?

94°C Signup and view all the answers

How long can denaturation of dsDNA take?

Up to 2 minutes Signup and view all the answers

What impact does a higher GC content have on denaturation temperatures?

Requires higher and longer denaturation temperatures Signup and view all the answers

What is the recommended annealing temperature (Ta) for PCR primers?

50-65 °C Signup and view all the answers

What happens if the annealing temperature in PCR is too high?

Primers do not anneal efficiently Signup and view all the answers

How long should the annealing time be maintained ideally in PCR?

15-60 seconds Signup and view all the answers

At what temperature can the extension step of PCR usually be carried out?

70-80°C for 1 to 2 minutes Signup and view all the answers

What is the optimal operating temperature for Taq polymerase during the extension step for the first 2 kb?

70-80°C for 1 minute Signup and view all the answers

How does the required time per additional kb differ between Taq and Pfu polymerases during the extension step?

Taq requires an additional 1 minute per kb, Pfu requires an additional 2 minutes per kb Signup and view all the answers

What is the main principle behind Gel Electrophoresis?

Separation and analysis of macromolecules based on size and charge Signup and view all the answers

What happens to charged biomolecules when placed in an electric field during Gel Electrophoresis?

They move toward the opposite charged electrode Signup and view all the answers

Which direction does DNA move in Gel Electrophoresis?

Toward the positive electrode Signup and view all the answers

What happens to negatively charged biomolecules in Gel Electrophoresis?

They move towards the anode (+) Signup and view all the answers

What happens to positively charged biomolecules in Gel Electrophoresis?

They move towards the cathode (-) Signup and view all the answers

How does size affect the movement of molecules in Gel Electrophoresis?

Larger molecules will be held up, smaller molecules will be impeded less and move faster Signup and view all the answers

What is the main role of the gel matrix in gel electrophoresis?

To serve as a porous medium through which molecules can move during electrophoresis Signup and view all the answers

What is the purpose of sample loading in gel electrophoresis?

To load samples containing the molecules to be separated into wells or slots at one end of the gel Signup and view all the answers

What is the function of the electrophoresis chamber in gel electrophoresis?

To provide an electric field for molecules within the gel to move toward the oppositely charged electrode Signup and view all the answers

What type of marker is commonly used to stain the gel after electrophoresis for DNA separation?

Ethidium bromide Signup and view all the answers

What information can be obtained by analyzing the separated molecules in the gel after electrophoresis?

Size, quantity, and purity of the molecules Signup and view all the answers

For what purpose is gel electrophoresis often used in relation to DNA fragments?

To determine the fragment size or check the success of DNA amplification Signup and view all the answers

Who should not take the new drug based on the SNP variation analysis?

Individuals not carrying the T allele on a specific locus in the CYP2D6 gene Signup and view all the answers

Which gene's variation analysis is relevant for determining who should not take the new drug?

CYP2D6 gene Signup and view all the answers

What did the SNP variation analysis reveal about the new drug's benefit for certain individuals?

It is not beneficial for individuals not carrying the T allele on a specific locus in the CYP2D6 gene Signup and view all the answers

What does the electrophoresis gel show?

The DNA of two male chimpanzees, a female, and an infant Signup and view all the answers

Why was the DNA from two male chimpanzees tested?

To determine the paternity of the infant chimpanzee Signup and view all the answers

What event triggered the DNA testing of the chimpanzees?

The recent birth of an infant by a female chimpanzee in captivity Signup and view all the answers

chapter 9

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Questions and Answers

What is the main purpose of Polymerase Chain Reaction (PCR)?

What does 'in vitro' refer to in the context of PCR?

How does PCR relate to DNA Replication in cells?

What is one of the applications of PCR mentioned in the text?

How does PCR contribute to forensic analysis?

In what way does PCR support drug development?

What is the purpose of DNA Extraction Lysis in sample preparation?

What is the role of Proteinase K in the DNA extraction process?

What is the final step in DNA purification during sample preparation?

What does PCR copy from the sample?

What information is required for the amplification of a specific target sequence using PCR?

What is the role of PCR primers in the amplification process?

What does PCR copy from the sample?

What information is required for the amplification of a specific target sequence using PCR?

What is the main purpose of Polymerase Chain Reaction (PCR)?

What is the purpose of oligonucleotide primers in PCR?

What is the role of DNA polymerase in PCR?

Why are two oligonucleotide primers required in PCR amplification?

Why are two primers required in a PCR reaction?

What is the role of the forward primer in PCR?

What happens if only one primer is used in a PCR reaction?

What is the recommended GC content range for a good primer?

Why is it important for the 3’ of a primer to end in G or C?

What impact do G and C bases have on the melting temperature (Tm) of primers?

At what temperature does denaturation of dsDNA usually occur?

How long can denaturation of dsDNA take?

What impact does a higher GC content have on denaturation temperatures?

What is the recommended annealing temperature (Ta) for PCR primers?

What happens if the annealing temperature in PCR is too high?

How long should the annealing time be maintained ideally in PCR?

At what temperature can the extension step of PCR usually be carried out?

What is the optimal operating temperature for Taq polymerase during the extension step for the first 2 kb?

How does the required time per additional kb differ between Taq and Pfu polymerases during the extension step?

What is the main principle behind Gel Electrophoresis?

What happens to charged biomolecules when placed in an electric field during Gel Electrophoresis?

Which direction does DNA move in Gel Electrophoresis?

What happens to negatively charged biomolecules in Gel Electrophoresis?

What happens to positively charged biomolecules in Gel Electrophoresis?

How does size affect the movement of molecules in Gel Electrophoresis?

What is the main role of the gel matrix in gel electrophoresis?

What is the purpose of sample loading in gel electrophoresis?

What is the function of the electrophoresis chamber in gel electrophoresis?

What type of marker is commonly used to stain the gel after electrophoresis for DNA separation?

What information can be obtained by analyzing the separated molecules in the gel after electrophoresis?

For what purpose is gel electrophoresis often used in relation to DNA fragments?

Who should not take the new drug based on the SNP variation analysis?

Which gene's variation analysis is relevant for determining who should not take the new drug?

What did the SNP variation analysis reveal about the new drug's benefit for certain individuals?

What does the electrophoresis gel show?

Why was the DNA from two male chimpanzees tested?

What event triggered the DNA testing of the chimpanzees?

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