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PCR stages

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Wize University Biology Textbook > DNA Replication & Repair

In Vitro DNA Replication: Polymerase Chain Reaction (PCR)

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Which is NOT a stage in the PCR cycle?
More In Vitro DNA Replication: Polymerase Chain Reaction (PCR) Questions:
PCR stages
Given the following DNA sequence, list the 6-base forward and reverse primers you can use to amplify the underlined region.
3'-AGCTGAGTATCGGTCCTAACAGTCAACCGCCT-5'
5'-TCGACTCATAGCCAGGATTGTCAGTTGGCGGA-3'
PCR stages
Which two primers will have the most similar melting temperatures (Tₘ)?
GCGATGT
ATGCGCA
In a PCR reaction, a region of the template strand reads 5′–GATTAC–3′ at the binding site for the reverse primer.
What is the sequence of the reverse primer in the 5' to 3' direction?
PCR is a method which has been used to...
You are studying a particular mutant fly which has misshapen wings. You believe you know which gene has been mutated to cause the misshapen wing phenotype but you do not know exactly what the mutation is (i.e. point mutation? Deletion? Insertion?) What is the best experimental protocol to determine the sequence of the mutated gene?
Suppose you start with one double-stranded DNA fragment which you then amplify by doing 25 rounds of PCR. How many DNA fragments would you have at the end of the procedure?
Cloning
You want to perform directed mutagenesis on a on a gene that is located in a plasmid. Following this change in amino acid sequence you want to over express the protein in a bacterial host for purification.
What enzymes, both inside and outside of the bacterial host, are necessary for this process?
Practice: DNA Techniques
You are studying a particular mutant fly which has misshapen wings. You believe you know which gene has been mutated to cause the misshapen wing phenotype but you do not know exactly what the mutation is (i.e. point mutation? Deletion? Insertion?) What is the best experimental protocol to determine the sequence of the mutated gene?
Primer annealing during PCR is an important step that is usually between 45-62 degrees Celsius and lasts for only 20-30 seconds. Which of the following statements best explains why the annealing time needs to be so short?
Why is a specific Taq Polymerase used in PCR reactions?
Suppose you start with one double-stranded DNA fragment which you then amplify by doing 25 rounds of PCR. How many DNA fragments would you have at the end of the procedure?
PCR is a method which has been used to...
You are studying a particular mutant fly which has misshapen wings. You believe you know which gene has been mutated to cause the misshapen wing phenotype but you do not know exactly what the mutation is (i.e. point mutation? Deletion? Insertion?) What is the best experimental protocol to determine the sequence of the mutated gene?
Why is a specific Taq Polymerase used in PCR reactions?
How does the polymerase chain reaction (PCR) bypass the need for a DNA helicase?
Which of the following is required for a PCR?
PCR is a method which has been used to.....
Suppose you start with one double stranded DNA fragment which you then amplify by doing 25 rounds of PCR. How many copies of the DNA fragments will you have by the end of the 25th round?
Which of the following is required for a PCR?
Match the process to the correct stage in PCR.