Essentials of Genetics (9th Edition) - Standalone book
9th Edition
ISBN: 9780134047799
Author: William S. Klug, Michael R. Cummings, Charlotte A. Spencer, Michael A. Palladino
Publisher: PEARSON
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Chapter 17, Problem 17PDQ
In a typical PCR reaction, describe what is happening in stages occurring at temperature ranges (a) 90–95°C, (b) 50–70°C, and (c) 70–75°C.
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In a typical PCR reaction what is happening in stages occurring at temperature ranges (a) 92–95°C, (b) 45–65°C, and (c) 65–75°C.
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.
Your 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: __________________________________
Chapter 17 Solutions
Essentials of Genetics (9th Edition) - Standalone book
Ch. 17 -
CASE STUDY |Should we worry about recombinant DNA...Ch. 17 - Prob. 2CSCh. 17 - Prob. 3CSCh. 17 -
HOW DO WE KNOW?
1. In this chapter we focused on...Ch. 17 - Prob. 2PDQCh. 17 - What roles do restriction enzymes, vectors, and...Ch. 17 - Prob. 4PDQCh. 17 - Prob. 5PDQCh. 17 - Prob. 6PDQCh. 17 - Prob. 7PDQ
Ch. 17 - List the advantages and disadvantages of using...Ch. 17 - What are the advantages of using a restriction...Ch. 17 - The introduction of genes into plants is a common...Ch. 17 - Prob. 11PDQCh. 17 - Prob. 12PDQCh. 17 - Prob. 13PDQCh. 17 - What advantages do cDNA libraries provide over...Ch. 17 - Prob. 15PDQCh. 17 -
16. List the steps involved in screening a...Ch. 17 -
17. In a typical PCR reaction, describe what is...Ch. 17 -
18. We usually think of enzymes as being most...Ch. 17 - How are dideoxynucleotides (ddNTPs) structurally...Ch. 17 - Prob. 20PDQCh. 17 - One complication of making a transgenic animal is...Ch. 17 -
22. When disrupting a mouse gene by knockout, why...Ch. 17 - Prob. 23PDQ
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- There 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_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_forwarda) (iii) (iv) Polymerase chain reaction (PCR) is a technique that enables multiplication of specific DNA sample at minute amount to millions or billions of copies at a short time span. (i) Figure 1 indicates the chemicals added into the PCR reaction tube prior to the addition of thermostable DNA polymerase. Do you agree with the list? Justify your answer. DNA template Buffer (containing Tris-HCI, KCI, Mg2+) ddNTP Forward primer Reverse primer Figure 1 If TA cloning is planned to be carried out after amplification of the gene, which thermostable DNA polymerase will you select and the reason for your selection? Why is the optimal annealing temperature vital in this technique? Explain how this temperature (too high or too low) will affect the efficiency of this reaction. If the primers you purchased possessed the following information: Number of Guanine: 4 Number of Adenine: 3 Number of Thymine: 4 Number of Cytosine: 5 Calculate the melting temperature of this primer and estimate the…arrow_forward
- A) What are the three main steps involved in PCR? Include temperatures and descriptions of each step. B) Explain why RFLP can produce many bands on an electrophoresis gel and PCR (one set of primers) will only produce one or two bands on a gel for the same genome.arrow_forwarda) The neomycin resistant colonies were screened by PCR using the primers A and B. Only seven of the 515 colonies (AB1 – AB7) resulted in the production of a PCR product of the expected size. The remaining 508 colonies failed to produce a PCR product (of any size). Explain why this would have been observed. Note: there is a rational explanation for this other than “the experiment did not work or there was a problem with the reagents”.arrow_forwardYou are working in a molecular biology laboratory and are having challenges with your PCR. You decide to double-check the PCR protocol programmed into the thermal cycler and discover that the annealing temperature was programmed to be 65 °C instead of 50 °C, as you had intended. What effects would this mistake have on the PCR reaction? Suggest potential solution(s).arrow_forward
- You have a 100 micromolar stock solution of dNTPs in the freezer and are setting up a PCR reaction in which you need to have a final concentration of 20 micromolar dNTPs. In a reaction volume of 25 microliters, how much of your stock dNTPs do you add to each PCR reaction?arrow_forward1) Prepare the following enzymatic reaction, present it in tabulated form. In a final volume of 30 ul, where buffer 4 (10 ml). How much volume of each reagent would be used and how much of water? Is there any problem? 2) The DNA pol 1 enzyme comes at a concentration of 50,000 U/ml. You have to prepare a 50 ug PCR reaction where you must use 0.05 U/ml reaction. You add 10 ul of PCR buffer, 2 ng of tempered DNA that is at a concentration of 0.5 ng/ul, primers (which are at 200 mM) so that each one remains at a concentration of 200 uM, Mg+2 that is 5 mM (10 X), enzyme and water. Present the table of all the reagents included in the reaction, the volumes of each one in ul. Present where the initial and final concentration of each reagent applies. Assume you have micropipettes for all values.arrow_forwardIn PCR amplification reaction, 5 mM EDTA was erroneously added to your final reaction, and you did not get any products. What is a possible explanation for this (based on the properties of EDTA)?arrow_forward
- You are setting up your PCR reaction and accidentally add twice as much of the salt buffer as you were supposed to. Select all that apply. 1. How will this impact product formation? 2. In what way(s) will the reaction be altered? (a) ...because primer/template binding will be altered (b) ...because the mechanism of dNTP addition will be altered. (c) You will get more of the desired PCR product... (d) ...because template denaturation will be altered. (e) You will get the same amount of the desired PCR product... (f) You will get less of the desired PCR product...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_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
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