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Questions and Answers
In PCR, what is the function of the thermostable DNA polymerase?
Why is Taq polymerase commonly used in PCR despite its known error introduction?
What are the two capabilities of thermostable DNA polymerases suitable for PCR?
At what temperature does the annealing step typically occur in a PCR cycle?
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What is the purpose of repeating the PCR reaction cycles for 30 or 40 cycles?
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Which enzyme is known for introducing errors in PCR due to its lack of 3' to 5' proofreading exonuclease activity?
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Which step of the PCR cycle involves the binding of primers to the DNA template?
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What is the half-life of Taq polymerase at 95°C, the denaturation temperature in PCR?
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Which enzyme, known for its greater accuracy, is used for certain PCR applications due to its 3' to 5' proofreading exonuclease activity?
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Which temperature is typically used for the annealing step in a PCR cycle?
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What is the function of the annealing step in a PCR cycle?
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What makes enzymes like Pfu DNA polymerase suitable for certain PCR applications?
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What is the role of the thermostable DNA polymerase in a PCR reaction cycle?
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Why is Taq polymerase commonly used in PCR despite its known error introduction?
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What is the purpose of repeating the PCR reaction cycles for 30 or 40 cycles?
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Study Notes
PCR and Thermostable DNA Polymerase
- Thermostable DNA polymerase is responsible for synthesizing new DNA strands during PCR by adding nucleotides to the primer sequences.
- Taq polymerase, a type of thermostable DNA polymerase, is commonly used in PCR despite introducing errors due to its lack of 3' to 5' proofreading exonuclease activity, because it is highly processive and can synthesize long strands of DNA.
Capabilities of Thermostable DNA Polymerases
- Thermostable DNA polymerases suitable for PCR have two capabilities: they can withstand high temperatures (necessary for DNA melting) and synthesize new DNA strands during the extension step.
PCR Cycle Steps
- The annealing step typically occurs at a temperature between 50°C to 65°C, allowing primers to bind to the DNA template.
- The annealing step involves the binding of primers to the DNA template.
- The purpose of repeating the PCR reaction cycles for 30 or 40 cycles is to amplify the target DNA sequence exponentially.
Enzyme Properties
- Taq polymerase has a half-life of 20-40 minutes at 95°C, the denaturation temperature in PCR.
- Pfu DNA polymerase, known for its greater accuracy, is used for certain PCR applications due to its 3' to 5' proofreading exonuclease activity, which corrects errors during DNA synthesis.
- Enzymes like Pfu DNA polymerase are suitable for certain PCR applications due to their high accuracy and error correction capabilities.
PCR Reaction Cycle
- The role of the thermostable DNA polymerase in a PCR reaction cycle is to synthesize new DNA strands during the extension step.
- The annealing step is necessary for primers to bind to the DNA template, allowing DNA synthesis to occur during the extension step.
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Description
Test your knowledge about Polymerase Chain Reaction (PCR) and its components, including denaturation, primer annealing, and polymerization. Learn about the steps involved in amplifying a sequence of DNA using PCR.
