BIOLOGY >PRINT UPGRADE<
11th Edition
ISBN: 9780357091586
Author: Solomon
Publisher: CENGAGE L
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Chapter 15.1, Problem 4LO
Compare the possible differences between a eukaryotic protein-encoding gene cloned by PCR and the same gene cloned by reverse transcriptase PCR (RT–PCR).
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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.
The exponential nature of PCR allows spectacular increases in the abundanceof a DNA sequence being amplified. Consider a 10-kbp DNA sequence in agenome of 1010 base pairs. What fraction of the genome does this sequence represent? That is, 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.
Explain why DNA ladders are usually included during gel electrophoresis.
One aspect of PCR that can be modified is the annealing temperature. In general, higher annealing temperatures show more specificity towards a single template, whereas lower annealing temperatures show less specificity and may bind to multiple regions throughout the genome. Discuss how using an annealing temperature that is too high or too low might influence the results of a PCR assay (and gel electrophoresis results) such as the one used in this study.
Chapter 15 Solutions
BIOLOGY >PRINT UPGRADE<
Ch. 15.1 - Prob. 1LOCh. 15.1 - Explain how gel electrophoresis is used to...Ch. 15.1 - Describe how PCR is used to amplify a specific...Ch. 15.1 - Compare the possible differences between a...Ch. 15.1 - Prob. 1CCh. 15.1 - Different forms of a protein are produced in the...Ch. 15.1 - What advantages does the PCR method have over gene...Ch. 15.2 - Describe the features of a typical CRISPR locus in...Ch. 15.2 - Explain the function of CRISPR in bacterial cells.Ch. 15.2 - Compare CRISPR-based endonucleases with...
Ch. 15.2 - Prob. 8LOCh. 15.2 - Prob. 1CCh. 15.2 - Prob. 2CCh. 15.2 - Prob. 3CCh. 15.3 - Prob. 9LOCh. 15.3 - Prob. 10LOCh. 15.3 - Discuss how qPCR, DNA microarrays (DNA chips), and...Ch. 15.3 - Explain how you would compare the expression of a...Ch. 15.3 - Prob. 2CCh. 15.4 - Describe how genome-wide association studies have...Ch. 15.4 - Explain how targeted gene silencing and knockout...Ch. 15.4 - Prob. 1CCh. 15.5 - Describe at least one important application of DNA...Ch. 15.5 - Prob. 1CCh. 15.5 - What are short tandem repeats (STRs), and why are...Ch. 15.5 - Why do gene targeting and mutagenesis screening in...Ch. 15.6 - Prob. 15LOCh. 15.6 - Prob. 16LOCh. 15.6 - Prob. 1CCh. 15.6 - Prob. 2CCh. 15.7 - Describe at least two safety issue associated with...Ch. 15.7 - What are some of the environment concerns...Ch. 15 - A plasmid (a) can be used as a DNA vector (b) is a...Ch. 15 - DNA molecules with complementary sticky ends...Ch. 15 - Prob. 3TYUCh. 15 - Which technique rapidly replicated specific DNA...Ch. 15 - Prob. 5TYUCh. 15 - A cDNA clone contains (a) introns (b) exons (c)...Ch. 15 - Prob. 7TYUCh. 15 - Gel electrophoresis separates nucleic acids on the...Ch. 15 - A CRISPR locus in a bacterium contains (a) short...Ch. 15 - DNA molecular with complementary sticky ends...Ch. 15 - These highly polymorphic molecular markers are...Ch. 15 - Prob. 12TYUCh. 15 - Prob. 13TYUCh. 15 - Prob. 14TYUCh. 15 - EVOLUTION LINK DNA technology, such as the...Ch. 15 - SCIENCE, TECHNOLOGY, AND SOCIETY What are some...
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- Polymerase Chain Reaction (PCR) was invented by Kary Mullis in 1983. This technique had indeed facilitated research in various areas of molecular biology and genetics. You would like to amplify a particular gene fragment from the yeast genome using Polymerase Chain Reaction (PCR). What are the THREE (3) main cycles in PCR? Discuss the processes at each PCR cycle mentioned.arrow_forwardExplain how the percentage efficiency of a real-time PCR reaction is determined using a theoretical experiment and why this is essential in any real-time PCR analysis.arrow_forwardPCR errors during library amplification are one possible source of false positive results. If an error occurs in the first round of amplification, all the subsequent copies of that library fragment will also carry the variant, even though it is not present in the genome. However, reads from other library fragments spanning this same region will not have the variant, which means that identifying PCR copies, or clones, of library fragments during analysis can help to identify these types of errors. Based on what you know about PCR, which of the following statements would be true about the PCR clones in a sequencing library? A. PCR is subject to amplification bias, so reads derived from PCR clones will only map to regions that are not GC-rich. B. PCR is subject to amplification bias, so reads derived from PCR clones will only map to non-repetitive regions. C. PCR produces identical copies, so reads derived from PCR clones would map to the exact same location. D. PCR introduces many…arrow_forward
- 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_forwardThe temperature at which the primers and target DNA hybridize may be changed to influence the stringency of PCR amplification. What effect will changing the hybridization temperature have on the amplification? Let's say you have a certain yeast gene A and want to check whether it has a human equivalent. How might managing the hybridization's rigor benefit you?arrow_forwardWhat are the different types of PCR and their product concepts? Site and explain the comparison briefly.arrow_forward
- Below 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_forwardWhat are the reasons why there needs to be more than 10 cycles in the PCR process? Name 3 reasons.arrow_forwardThere are many types of PCR techniques available nowadays. Explain the concept of nested PCR and real-time PCR.arrow_forward
- Evaluation of PCR product electrophoresed on 0.8% agarose gel shows non-specific bands. The appropriate modification for the next PCR reaction is to: a) increase the concentration of Taq polymerase b) increase the number of PCR cycles c) decrease the template denaturation temperature d) reduce the concentration of primersarrow_forwardWhat are the reagents used for Reverse Transcriptase PCR (RT-PCR) and Reverse Transcriptase–Real time–PCR (RT-qPCR)?arrow_forwardMention all the enzymes and reagents used in the everse Transcriptase Polymerase Chain Reaction (RT-PCR) technique, with their functions.arrow_forward
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