Brock Biology of Microorganisms (15th Edition)
15th Edition
ISBN: 9780134261928
Author: Michael T. Madigan, Kelly S. Bender, Daniel H. Buckley, W. Matthew Sattley, David A. Stahl
Publisher: PEARSON
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Textbook Question
Chapter 19.6, Problem 1MQ
- What could you conclude from PCR/DGGE analysis of a sample that yielded one band by PCR and one band by DGGE? One band by PCR and four bands by DGGE?
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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?
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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 answer
What are the reasons why there needs to be more than 10 cycles in the PCR process? Name 2 reasons.
Chapter 19 Solutions
Brock Biology of Microorganisms (15th Edition)
Ch. 19.1 - Describe the enrichment strategy behind...Ch. 19.1 - Why is sulfate (So42) added to a Winogradsky...Ch. 19.1 - What is enrichment bias? How does dilution reduce...Ch. 19.1 - Why do the results of a direct enrichment of an...Ch. 19.2 - What is a pure culture and why is obtaining one...Ch. 19.2 - How does the agar dilution method differ from...Ch. 19.2 - What criteria serve to demonstrate that a culture...Ch. 19.3 - How might you isolate a morphologically unique...Ch. 19.3 - What is meant by high-throughput in culturing...Ch. 19.3 - What feature of high-throughput culturing relieves...
Ch. 19.4 - How does viability staining differ from stains...Ch. 19.4 - What types of environments limit the application...Ch. 19.4 - Why is it incorrect to say that the GFP is a...Ch. 19.4 - Prob. 1CRCh. 19.5 - What structure in the cell is the target for...Ch. 19.5 - FISH and CARD-FISH can be used to reveal different...Ch. 19.5 - Why is CARD-FISH more suitable than FISH for...Ch. 19.6 - What could you conclude from PCR/DGGE analysis of...Ch. 19.6 - What surprising finding has come out of many...Ch. 19.6 - How has next-generation sequencing technology...Ch. 19.6 - QWhich method, ARISA or T-RFLP, would provide more...Ch. 19.7 - Prob. 1MQCh. 19.7 - What are the advantages and disadvantages of...Ch. 19.7 - Why might a microarray be superior to using...Ch. 19.8 - Prob. 1MQCh. 19.8 - How do environmental genomic approaches differ...Ch. 19.8 - Prob. 3MQCh. 19.8 - Prob. 1CRCh. 19.9 - Prob. 1MQCh. 19.9 - If a large pulse of organic matter entered the...Ch. 19.9 - Q What are the major advantages of radioisotopic...Ch. 19.10 - What is the simplest explanation for why lunar...Ch. 19.10 - What is the expected isotopic composition of...Ch. 19.10 - How might exchange of metabolites among members of...Ch. 19.10 - Will autotrophic organisms contain more or less...Ch. 19.11 - How could NanoSIMS be used to identify a...Ch. 19.11 - Prob. 2MQCh. 19.11 - How does MAR-FISH link microbial diversity and...Ch. 19.11 - Q What can MAR-FISH tell you that FISH alone...Ch. 19.12 - How can stable isotope probing reveal the identity...Ch. 19.12 - What key method is required to do genomics on a...Ch. 19.12 - Prob. 3MQCh. 19.12 - How would you use cytometric cell sorting to...Ch. 19 - Design an experiment for measuring the activity of...Ch. 19 - You wish to know whether Archaea exist in a lake...Ch. 19 - Design an experiment to solve the following...Ch. 19 - Design a SIP experiment that would allow you to...
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- Explain 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_forwardYou have added 4 μl of a 5 μM stock primer to a 25 μl PCR reaction. What is the amountof primer in nanomol that you have added to the reaction?arrow_forwardThe 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.arrow_forward
- a) 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_forwarda) Which step of RT-PCR reactions does the figure shown below represents? b) Describe all enzyme(s) and name the molecule(s) that take part or are products of this step? Enzymes: Molecule 1: Molecule 2: Molecule 3: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_forward
- Choose the correct statements from the list below. There may be more than one correct statement. A) If you start with 2 DNA templates, after four rounds of PCR you'll have 32 copies B) PCR is useful in making millions or billions of copies of a gene so that it is present in a quantity large enough to study C) quantitative PCR is very similar to PCR, but fluorescent probes are added so that we can measure how much PCR product exists by examining how much the reaction fluoresces D) In real-time reverse transcriptase PCR, the RNA is used as a template to make a cDNA copy (through reverse transcriptase)arrow_forwardA region of the genome is amplified by PCR to analyze two closely linked SNPs. PCR products from individual A were labelled with a red-fluorescing dye and those from individual B labelled with a green-fluorescing dye. These PCR products were mixed and hybridized to an oligonucleotide microarray as shown in the figure below. What can we conclude about the data shown? Select all true: a. individual A is heterozygous for the M1 and M2 haplotypes, individual B is heterozygous for M4 and M5 b. individual A is heterozygous for the M1 and M2 haplotypes, individual B is heterozygous for M3 and M6 c. Sanger sequencing traces of these PCR products would show the same double peaks at the same positions for these two individuals d. analyzing this PCR amplified region with conventional Sanger sequencing would be more accurate than the microarray analysis e. individuals A and B could be siblings who share a parent that is homozygous for one of the haplotypes {urgent}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
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