10. For each of the following reagents briefly explain their role in a PCR reaction (3...
10. For each of the following reagents briefly explain their role in a PCR reaction (3 points each) a. dNTP b. DNA containing the sequence to be amplified. c. DNA ligase. d. Heat-stable DNA polymerase. e. Oligonucleotide primer(s).
Homework Answers
DNTP- THESE are nucleotide required in DNA synthesis in reaction. They are monomer of DNA.
DNA CONTAINING SEQUENCE TO BE AMPLIFIED - it is called template at which reaction occur and this region is amplified in many copies.
DNA ligase - it join phosphodiester bonds in DNA.
HEAT STABLE DNA POLYMERASE - As very high temperature is used in this reaction, so polymerase should be heat stable.
Oligonucleotide primer - primer is required in PCR as Taq polymerase can't initiate reaction itself.
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10. For each of the following reagents briefly explain their
role in a PCR reaction (3...
2. You are performing PCR under the following conditions: 1. Each PCR reaction contains 50 pl...
2. You are performing PCR under the following conditions: 1. Each PCR reaction contains 50 pl final volume II. Each PCR reaction contains: 1x PCR buffer, 1.5 mM MgCl2, 0.2 mM dNTP mix, 0.2 MM sense primer, 0.2 uM anti-sense primer, 1.5 units of Taq polymerase and 2 ul of DNA template. [all concentrations are final] III. Stock solutions of the following reagents are available: 10x PCR buffer, 25 mm MgCl2, 10 mM dNTP mix, 10 M of sense primer,...
Question 33 (2 points) The requirements of the polymerase chain reaction (PCR) include: a) knowing parts...
Question 33 (2 points) The requirements of the polymerase chain reaction (PCR) include: a) knowing parts of a target DNA sequence to be amplified. b) two primers, complementary to each end of the target sequence. c) a large supply of DNA nucleotides. d) a heat-stable DNA polymerase. e) All of the above
Which of the following statements about the polymerase chain reaction (PCR) is false? a The DNA...
Which of the following statements about the polymerase chain reaction (PCR) is false? a The DNA ligase used is from a thermophilic bacteria so that it can resist denaturation at high temperatures. b Newly synthesized DNA must be heat-denatured before the next round of DNA synthesis begins. c The boundaries of the amplified DNA segment are determined by the synthetic oligonucleotides used to prime DNA synthesis. d DNA amplified by PCR can be cloned.
CALCULATIONS [2 points each] You are asked to perform a PCR experiment in the lab. However, in th...
CALCULATIONS [2 points each] You are asked to perform a PCR experiment in the lab. However, in this experiment you must add each ingredient separately (unlike the "master-mix" we used in our own experiment). In this experiment, each PCR reaction tube contained the following ingredients added with a micropipetor... 10 HL of 5x Buffer 6 μ1 of 25 mM MgCl2 solution μL of 10 mM dNTP mix 1.5 μL oligonucleotide primer mix 0.5 HL Taq DNA Polymerase stock solution 28...
25. Which is not a required reactant for a PCR reaction? Select one: a. Primer DNA...
25. Which is not a required reactant for a PCR reaction? Select one: a. Primer DNA b. Taq polymerase c. dNTP d. template DNA e. ATP
Which of the following is needed for the PCR technique to work? (multiple answers) a. A...
Which of the following is needed for the PCR technique to work? (multiple answers) a. A probe that can anneal to the center of the target DNA sequence. b. A thermostable RNA polymerase c. Large amounts of pure DNA d. A thermostable DNA polymerase e. Two DNA primers that can anneal to the target DNA on either side of the DNA to be amplified. f. A thermostable DNA ligase
Which of the following components is NOT used in the technique of PCR to amplify a...
Which of the following components is NOT used in the technique of PCR to amplify a specific DNA sequence in a mouse genome? O Specific single-stranded DNA primers O Double-stranded genomic mouse DNA O Heat-stable DNA polymerase D DNA ligase
1.The PCR (polymerase chain reaction) protocol that is currently used in laboratories was facilitated by the...
1.The PCR (polymerase chain reaction) protocol that is currently
used in laboratories was facilitated by the discovery of a
bacterium called Thermus aquaticus in a hot spring inside
Yellowstone National Park, in Wyoming. This organism contains a
heat-stable form of DNA polymerase known as Taq
polymerase, which continues to function even after it has been
heated to 95°C.
a.Why would such a heat-stable polymerase be beneficial in
PCR?
b.What would happen if it weren’t
heat stable?
c.How might you choose...
What is the purpose in a PCR reaction for each of the following reagents? Taq Taq...
What is the purpose in a PCR reaction for each of the following reagents? Taq Taq buffer dNTPs Forward primer Reverse primer Genomic DNA What do you think would happen if you forgot to add your Reverse primer when you did a PCR?
Now. you should be able to answer the following questions: • How the amplification will be...
Now. you should be able to answer the following questions: • How the amplification will be done? - How you will determine your target sequence? How the amplification will be specific for certain segment? What are the requirements to carry PCR? • Suppose you perform a PCR that begins with one double-strand of the following DNA template: +5'-CTACCTGCGGGTTGACTGCTACCTTCCCGGGATGCCCAAAATTCTCGAG-3+ +3'-GATGGACGCCCAACTGACGATGGAAGGGCCCTACGGGTTTTAAGAGCTC-5'+ A. Draw one cycle of PCR reaction below the following diagram. B. Label the template DNA, the primers, and what is...
2. You are performing PCR under the following conditions: 1. Each PCR reaction contains 50 pl final volume II. Each PCR reaction contains: 1x PCR buffer, 1.5 mM MgCl2, 0.2 mM dNTP mix, 0.2 MM sense primer, 0.2 uM anti-sense primer, 1.5 units of Taq polymerase and 2 ul of DNA template. [all concentrations are final] III. Stock solutions of the following reagents are available: 10x PCR buffer, 25 mm MgCl2, 10 mM dNTP mix, 10 M of sense primer,...
Question 33 (2 points) The requirements of the polymerase chain reaction (PCR) include: a) knowing parts of a target DNA sequence to be amplified. b) two primers, complementary to each end of the target sequence. c) a large supply of DNA nucleotides. d) a heat-stable DNA polymerase. e) All of the above
CALCULATIONS [2 points each] You are asked to perform a PCR experiment in the lab. However, in this experiment you must add each ingredient separately (unlike the "master-mix" we used in our own experiment). In this experiment, each PCR reaction tube contained the following ingredients added with a micropipetor... 10 HL of 5x Buffer 6 μ1 of 25 mM MgCl2 solution μL of 10 mM dNTP mix 1.5 μL oligonucleotide primer mix 0.5 HL Taq DNA Polymerase stock solution 28...
Which of the following components is NOT used in the technique of PCR to amplify a specific DNA sequence in a mouse genome? O Specific single-stranded DNA primers O Double-stranded genomic mouse DNA O Heat-stable DNA polymerase D DNA ligase
1.The PCR (polymerase chain reaction) protocol that is currently
used in laboratories was facilitated by the discovery of a
bacterium called Thermus aquaticus in a hot spring inside
Yellowstone National Park, in Wyoming. This organism contains a
heat-stable form of DNA polymerase known as Taq
polymerase, which continues to function even after it has been
heated to 95°C.
a.Why would such a heat-stable polymerase be beneficial in
PCR?
b.What would happen if it weren’t
heat stable?
c.How might you choose...
Now. you should be able to answer the following questions: • How the amplification will be done? - How you will determine your target sequence? How the amplification will be specific for certain segment? What are the requirements to carry PCR? • Suppose you perform a PCR that begins with one double-strand of the following DNA template: +5'-CTACCTGCGGGTTGACTGCTACCTTCCCGGGATGCCCAAAATTCTCGAG-3+ +3'-GATGGACGCCCAACTGACGATGGAAGGGCCCTACGGGTTTTAAGAGCTC-5'+ A. Draw one cycle of PCR reaction below the following diagram. B. Label the template DNA, the primers, and what is...
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