Genetics: Analysis and Principles
5th Edition
ISBN: 9780073525341
Author: Robert J. Brooker Professor Dr.
Publisher: McGraw-Hill Education
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Chapter 20.2, Problem 1COMQ
Summary Introduction
Introduction:
The PCR (polymerase chain reaction) is the technique that is used to producemany copies of the specific DNA (deoxyribonucleic acid). This technique is based on the use of DNA polymerase enzyme.
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Below are several problems frequently faced by researchers when running the PCR. Give one
(1) solution to each problem stated.
1. No PCR product
2. Multiple bands appeared after gel electrophoresis (you are amplifying just 1
gene)
3. Bright thick bands at the end of the agarose gel after electrophoresis
4. Bands appeared at the negative control
Which of the following statements about PCR or RT-gPCR is correct?
1. Fluorescence is detected when the TagMan probe is cleaved by Taq polymerase (AMPLITAQ).
2. PCR uses the natural process of transcription to amplify DNA.
3. Both PCR and RT-qPCR require forward and reverse primers.
4. Fluorescence is detected when the reporter dye is in close proximity to the quencher dye.
A. 1, 2 and 3
B. 1 and 3
C. 2 and 4
D. 4 only
E. All of 1, 2, 3 and 4 are correct
Match the following terms with their definitions and label each component of the PCR mixture in the diagram (use the letters A-D):I. DNA polymeraseII. PrimersIII. NucleotidesIV. Genomic DNA template
A. DNA that contains the target sequence that will be replicated using PCR.B. An enzyme that copies the DNA sequence.C. A mixture of 4 nucleotides (A,G,C, and T) that will be polymerized into the replicated DNA sequence.D. A short DNA sequence that allows the enzyme to bind and initiate polymerization.
Chapter 20 Solutions
Genetics: Analysis and Principles
Ch. 20.1 - 1. Which of the following may be used as a vector...Ch. 20.1 - The restriction enzymes used in gene-cloning...Ch. 20.1 - 3. Which is the proper order of the following...Ch. 20.1 - 4. The function of reverse transcriptase is...Ch. 20.1 - A collection of recombinant vectors that carry...Ch. 20.2 - Prob. 1COMQCh. 20.2 - Prob. 2COMQCh. 20.2 - 3. During real-time PCR, the synthesis of PCR...Ch. 20.3 - When a dideoxyribonucleotide is incorporated into...Ch. 20.4 - 1. The purpose of site-directed mutagenesis and...
Ch. 20.5 - Which of the following methods use(s) a labeled...Ch. 20.5 - 2. Which of the following methods is used to...Ch. 20.5 - During Western blotting, the primary antibody...Ch. 20.6 - 1. In an EMSA, the binding of a protein to...Ch. 20.6 - The basis for DNase I footprinting is that the...Ch. 20 - Discuss three important advances that have...Ch. 20 - Prob. 2CONQCh. 20 - Write a double-stranded DNA sequence that is 20...Ch. 20 - What is cDNA? In eukaryotes, how does cDNA differ...Ch. 20 - 5. Draw the structural feature of a...Ch. 20 - Prob. 1EQCh. 20 - Prob. 2EQCh. 20 - Describe the important features of cloning...Ch. 20 - 4. How does gene cloning produce many copies of a...Ch. 20 - Prob. 5EQCh. 20 - Prob. 6EQCh. 20 - Prob. 7EQCh. 20 - Prob. 8EQCh. 20 - Prob. 9EQCh. 20 - Starting with a sample of RNA that contains the...Ch. 20 - 11. What type of probe is used for real-time PCR?...Ch. 20 - 12. What phase of PCR (exponential, linear, or...Ch. 20 - 13. DNA sequencing can help us to identify...Ch. 20 - A sample of DNA was subjected to automated DNA...Ch. 20 - Prob. 15EQCh. 20 - Prob. 16EQCh. 20 - Prob. 17EQCh. 20 - Prob. 18EQCh. 20 - Prob. 19EQCh. 20 - E20. In Southern, Northern, and Western blotting,...Ch. 20 - What is the purpose of a Northern blotting...Ch. 20 - Prob. 22EQCh. 20 - Prob. 23EQCh. 20 - 23. In the Western blot shown here, proteins were...Ch. 20 - If you wanted to know if a protein was made during...Ch. 20 - Prob. 27EQCh. 20 - Prob. 29EQCh. 20 - Prob. 31EQCh. 20 - 28. Describe the rationale behind the...Ch. 20 - Certain hormones, such as epinephrine, can...Ch. 20 - An electrophoretic mobility shift assay can be...Ch. 20 - Prob. 35EQCh. 20 - Prob. 36EQCh. 20 - Prob. 37EQCh. 20 - Prob. 24EQCh. 20 - Prob. 30EQCh. 20 - Prob. 25EQCh. 20 - Prob. 1QSDC
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- Each PCR cycle has three steps: DNA sample denaturation, primer annealing, and elongation/extension of the target DNA. Each cycle can theoretically double the original amount of DNA if the efficiency is 100%. True Falsearrow_forwardWill any of the PCR reaction components (e.g., dNTPs, primer) become limiting before the product is formed?arrow_forwardThe three steps involved in PCR are: Question 4 options: Denaturation, primer annealing, and extension Denaturation, ligation, and extension Denaturation, template annealing, and extension Denaturation, primer annealing, and exhaustionarrow_forward
- Look at each PCR component listed below. For each one, determine which steps(s) of the PCR reaction (denaturation, annealing or extension) would be directly affect if that component were missing. Taq polymerase: Oligonucleotide primers: DNA template: Deoxynucleotides (A, T, G and C): Imagine that you correctly prepare your PCR reaction mixture, but there is something wrong with the thermal cycler. Describe what would happen if: The thermal cycler was stuck on 940C: The thermal cycler cycled between 600C and 720C, but never reached 940C The thermal cycler cycled between 940C and 720C, but never reached 600arrow_forwardDescribe the three steps of a PCR cycle.arrow_forwardDefine the term PCR.arrow_forward
- All of the following are performed during restriction fragment length polymorphism analysis. 1. splitting of double-stranded into single-stranded DNA 2. gel electrophoresis 3. autoradiography 4. immersion in radioactive probes 5. digestion of DNA with restriction endonucleases 6. use of a positive charge to transfer single-stranded DNA from a gel to a membrane. The correct sequence of these operations is whatarrow_forwardLink the technique to the correct enzyme. PCR cutting DNA cDNA synthesis DNA sequencing pasting DNA a. reverse transcriptase b. Taq polymerase c. DNA polymerase (not Taq) d. DNA ligase e. restriction enzymearrow_forwardIf the RT-PCR test is negative, give instances when repeat RT-PCR test should be done. If repeated, how many days after the first test should the 2nd test be done?arrow_forward
- During a PCR reaction, all the following events occur except: A. Hybridization B. Binding of a 3’OH group in the active site of DNA polymerase C. Addition of nucleotide monomers on the 3’ end of the template DNA D. DNA denaturation E. Semi conservative replicationarrow_forwardExplain why a positive control and negative control are included in PCR experiments. Explain the three steps involved in each cycle of polymerase chain reaction.Why is loading dye added to the DNA sample for gel electrophoresis? Explain the function of the following components in a PCR reaction:− Primer, dNTP, MgCl, Taq polymerase, buffer.arrow_forwardPrimer annealing is an important aspect of PCR. The annealing step of the cycle usually takes place at around 45-55°C and lasts for only 20-30 seconds. Which of the following statements best explains why the annealing time has to be so brief? OA. A longer annealing time would prevent primers binding to specific sites. OB. The short time frame prevents primers from binding to each other. OC. If the time were any longer, DNA polymerase would begin to denature. OD. The short time frame minimizes complementary template strands base pairing with each other. Reset Selectionarrow_forward
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