Genetics: Analysis and Principles
6th Edition
ISBN: 9781259616020
Author: Robert J. Brooker Professor Dr.
Publisher: McGraw-Hill Education
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Textbook Question
Chapter 21, Problem 12EQ
What phase of PCR (exponential, linear, or stationary) is analyzed to quantitate the amount of DNA or RNA in a sample? Explain why this phase is chosen.
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The final amount of each primer required in a PCR reaction is 25 picomol. If the total volume of the PCR reaction is equal to 100 µl and the stock concentration of each primer is equal to 0.0025 mM. Calculate the volume of stock primer that needs to be added in order to ensure a primer amount of 25 picomol.
In a typical PCR reaction, describe what is happening in stages occurring at temperature ranges (a) 92–95°C, (b) 45–65°C, and (c) 65–75°C.
What phase of PCR (exponential, linear, or stationary) is analyzedto quantitate the amount of DNA or RNA in a sample? Explainwhy this phase is chosen.
Chapter 21 Solutions
Genetics: Analysis and Principles
Ch. 21.1 - 1. Which of the following may be used as a vector...Ch. 21.1 - The restriction enzymes used in gene-cloning...Ch. 21.1 - 3. Which is the proper order of the following...Ch. 21.1 - 4. The function of reverse transcriptase is...Ch. 21.1 - A collection of recombinant vectors that carry...Ch. 21.2 - Prob. 1COMQCh. 21.2 - Prob. 2COMQCh. 21.2 - 3. During real-time PCR, the synthesis of PCR...Ch. 21.3 - When a dideoxyribonucleotide is incorporated into...Ch. 21.4 - 1. The purpose of site-directed mutagenesis and...
Ch. 21.5 - Which of the following methods use(s) a labeled...Ch. 21.5 - 2. Which of the following methods is used to...Ch. 21.5 - During Western blotting, the primary antibody...Ch. 21.6 - 1. In an EMSA, the binding of a protein to...Ch. 21.6 - The basis for DNase I footprinting is that the...Ch. 21 - Discuss three important advances that have...Ch. 21 - Prob. 2CONQCh. 21 - Write a double-stranded DNA sequence that is 20...Ch. 21 - What is cDNA? In eukaryotes, how does cDNA differ...Ch. 21 - 5. Draw the structural feature of a...Ch. 21 - Prob. 1EQCh. 21 - Prob. 2EQCh. 21 - Describe the important features of cloning...Ch. 21 - 4. How does gene cloning produce many copies of a...Ch. 21 - Prob. 5EQCh. 21 - Prob. 6EQCh. 21 - Prob. 7EQCh. 21 - Prob. 8EQCh. 21 - Prob. 9EQCh. 21 - Starting with a sample of RNA that contains the...Ch. 21 - 11. What type of probe is used for real-time PCR?...Ch. 21 - 12. What phase of PCR (exponential, linear, or...Ch. 21 - 13. DNA sequencing can help us to identify...Ch. 21 - A sample of DNA was subjected to automated DNA...Ch. 21 - Prob. 15EQCh. 21 - Prob. 16EQCh. 21 - Prob. 17EQCh. 21 - Prob. 18EQCh. 21 - Prob. 19EQCh. 21 - What is the purpose of a Northern blotting...Ch. 21 - Prob. 21EQCh. 21 - Prob. 22EQCh. 21 - 23. In the Western blot shown here, proteins were...Ch. 21 - If you wanted to know if a protein was made during...Ch. 21 - Prob. 25EQCh. 21 - Prob. 26EQCh. 21 - Prob. 27EQCh. 21 - 28. Describe the rationale behind the...Ch. 21 - Certain hormones, such as epinephrine, can...Ch. 21 - An electrophoretic mobility shift assay can be...Ch. 21 - Prob. 31EQCh. 21 - Prob. 32EQCh. 21 - Prob. 33EQCh. 21 - Prob. 1QSDC
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- What would be the final primer concentration if 0.5 μl of 10 μM primers were added to a PCR reaction with a final volume of 20 μl?arrow_forwardWhat determines the elongation (extension) reaction time in pcr reaction?arrow_forwardBelow are gel electrophoresis results for initial and nested PCR of the Shrimp Plant. Use the results to answer these questions: a) Why does the Arabidopsis control generate two PCR bands, but the plasmid control only generates one PCR band? b) Did the negative control generate a PCR product? If so, what are the implications of this for the experiment? c) Would you use this nested PCR product from Shrimp Plant for cloning? Briefly explain your answerarrow_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_forwardYour amplicon from PCR was subjected to AGE for analysis. You are shown the image of the gel loaded with the following lanes: (A) negative control, (B) size ladder (1, 2, 3, 4, 6, and 10 kb), (C) gDNA extract, (D) PCR amplicon. However, due to mishaps while loading the gel with the samples, you are not sure which lane is which. You are shown a diagram of the obtained gel below. a. Label each lane of the gel. Write only the corresponding letters in the wells above. b. Above each band in the size ladder, write its size (in kb). c. Approximate the size (in kb) of the polystyrenase gene. Write your answer above the band corresponding to the gene. Bonus: If you wish to identify the bacterial species in this scenario, what gene is most commonly and routinely sequenced? Answer: __________________________________arrow_forwardWhat determines the size (length) of the primary PCR product? What might a successful gel check of a PCR reaction look like?arrow_forward
- The exponential nature of PCR allows spectacular increases in the abundance of a DNA sequence being amplified. Consider a 10-kbp DNA sequence in a genome of 1010 base pairs. What fraction of the genome is represented by this sequence; i.e., what is the fractional abundance of this sequence in this genome? Calculate the fractional abundance of this target sequence after 10, 15, and 20 cycles of PCR, starting with DNA representing the whole genome and assuming that no other sequences in the genome undergo amplification in the process.arrow_forwardOur PCR samples already contain loading dye, but sometimes this isn’t the case. If your samples didn’t already contain dye and you wanted to load your PCR sample onto an agarose gel, you’d need to add loading dye to the proper concentration. There is a 6X loading dye available for use; how many µl of this loading dye will you add to 10 µl of your sample so that it is at a 1X working concentration? Show your work.arrow_forwardThere are many PCR techniques available to suit the needs of all researchers in their laboratory task. (i) (ii) What is the major difference in the functions performed by the conventional PCR and real time PCR? Shania is planning to study the gene expression of Escherichia coli after a drug- treatment. She needs to decide between two types of chemistries to detect her PCR products (TaqMan Chemistry vs. SYBR Chemistry) using real-time PCR instruments. Compare and contrast between TaqMan Chemistry and SYBR Chemistry.arrow_forward
- let three loci be X,Y,Z. three pairs of primes can be used to amplify these loci in multiplex PCR. the resulting amplicons(from different loci) are of the equal size. will this scenario be acceptable in STR typing? why or why not?arrow_forwardYou want to set up 50 µl total volume of a PCR reaction. You have a microfuge tube of Taq polymerase (5 units/ul), and each PCR reaction requires a final of 10 units of Taq polymerase. You have a microfuge tube of Taq Buffer (5X), and each PCR reaction requires a final of 1X Taq Buffer. How much of Taq polymerase and Taq buffer would you add? Select all that apply 10 ul of Taq buffer 2 ul of Taq polymerase 5 ul of Taq buffer 5 ul of Taq polymerasearrow_forwardWhat would be the expected effect on the PCR reaction, if you increased the temperature of the annealing phase and the length of the elongation phase? O Accuracy will be reduced, but yield will be increased Accuracy and yield will be increased OAccuracy will be increased, but yield will be decreased Accuracy and yield will be reducedarrow_forward
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