la: The 2x PCR Master mix has four main components. Two of these are PCR Buffer and MgC12. Based on your knowledge of DNA replication and PCR, what are the other two components and what are their functions? 1b: Explain what would happen if the PCR reaction did not contain Taq polymerase? Assuming all other required PCR components are present, would any part of the PCR reaction occur? Explain your reasoning. 1c: Why does a PCR reaction require a primer? Would you expect this primer to be composed of DNA or RNA? Explain your reasoning.

Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
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Chapter1: Biochemistry: An Evolving Science
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PCR Analysis
la: The 2x PCR Master mix has four main components. Two of these are PCR Buffer and
MgCl2. Based on your knowledge of DNA replication and PCR, what are the other two
components and what are their functions?
1b: Explain what would happen if the PCR reaction did not contain Taq polymerase?
Assuming all other required PCR components are present, would any part of the PCR
reaction occur? Explain your reasoning.
1c: Why does a PCR reaction require a primer? Would you expect this primer to be
composed of DNA or RNA? Explain your reasoning.
1d: Should PCR primers be complementary to each other? Explain your reasoning
le: As you know, experiments include controls. In this lab, a negative control was included
along with your experimental samples. Which components do you think the negative control
would contain?
Hint: Think about which component would be left out of the negative control relative to the
experimental samples. Explain why.
1f: What would the PCR product be for the following DNA sequence at the end of one cycle
of amplification? Indicate the polarities of the product in your answer.
NOTE: This sequence does not start with AUG
3' - ACC CGA TAC GGA GTC AAA TTT – 5'
Transcribed Image Text:PCR Analysis la: The 2x PCR Master mix has four main components. Two of these are PCR Buffer and MgCl2. Based on your knowledge of DNA replication and PCR, what are the other two components and what are their functions? 1b: Explain what would happen if the PCR reaction did not contain Taq polymerase? Assuming all other required PCR components are present, would any part of the PCR reaction occur? Explain your reasoning. 1c: Why does a PCR reaction require a primer? Would you expect this primer to be composed of DNA or RNA? Explain your reasoning. 1d: Should PCR primers be complementary to each other? Explain your reasoning le: As you know, experiments include controls. In this lab, a negative control was included along with your experimental samples. Which components do you think the negative control would contain? Hint: Think about which component would be left out of the negative control relative to the experimental samples. Explain why. 1f: What would the PCR product be for the following DNA sequence at the end of one cycle of amplification? Indicate the polarities of the product in your answer. NOTE: This sequence does not start with AUG 3' - ACC CGA TAC GGA GTC AAA TTT – 5'
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