Life: The Science of Biology
11th Edition
ISBN: 9781319010164
Author: David E. Sadava, David M. Hillis, H. Craig Heller, Sally D. Hacker
Publisher: W. H. Freeman
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Chapter 13.5, Problem 2R
Summary Introduction
To review:
The basis of PCR test.
Introduction:
PCR stands for polymerase chain reaction that is an enzymatic method that helps in detecting certain genes in a pool of DNA (deoxyribonucleic acid) mix. It uses primers to make a complementary strand of the existing DNA molecule.
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What is needed from the cells for PCR?
What are the different types of PCR and their application? Please answer at your own words.
List the ingredients and describe how a PCR reaction works
Chapter 13 Solutions
Life: The Science of Biology
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- Why different temperatures are used in a PCR reaction? Please answer at your own words.arrow_forwardthey decided to check which bacterial colonies contains the recombinant gene W by PCR. they designed one set of primers, gene W forward, and gene W reverse, to amplify gene W. Complete the PCR profile that will be used below.arrow_forwardIf you wanted to prepare only one PCR reaction, how much of each reagent would you add to the PCR tube?arrow_forward
- In the second step of the PCR, the temperature is reduced to between 45-65 degrees C. What happens during this step?arrow_forwardwhat are ways you can run a PCR cycle in a single test tube? how would you adjust amplificons, enzymes, ddNTPs, Mg concentration so that it would be successful?arrow_forwardDescribe the three steps of a PCR cycle.arrow_forward
- Why is it important to wear gloves when setting up the PCR tubes?arrow_forwardA urine sample has been obtained, and the bacteria in this sample were cultured. To obtain more information regarding the identity of this Gram-negative strain, Sanger sequencing can be used. A bacterial colony is transferred into a 0.2 mL tube containing buffer, then boiled to break open the bacterial cells. The tube is centrifuged, and some of the supernatant is transferred to a PCR tube. Next, the following reagents are added: DNA polymerase, a primer that binds near the 16S rRNA region of the bacterial chromosome, dNTPs, and fluorescently-labeled ddNTPs. The sequencing reaction is processed in a thermocycler, then analyzed by capillary electrophoresis. This experiment generates the following results (in FASTA format): > sequencing results TAACAGGAAGCAGCTTGCTGCTTTGCTGACGAGTGGCGGACGGGTGAGTAATG TCTGGGAAACTGCCTGATGGAGGGGGATAACTACTGGAAACGGTAGCTAATAC CGCATAACGTCGCAAGCACAAAGAGGGGGACCTTAGGGCCTCTTGCCATCGGA TGTGCCCAGATGGGATTAGCTAGTAGGTGGGGTAACGGCTCACCTAGGCGACG…arrow_forwardIf the PCR analysis of your sample did not work can you suggest why? How can you test if this is true?arrow_forward
- What is a PCR?arrow_forwardWhich is a complete list of the ingredients that are essential for PCR? nucleotides, DNA template, Taq polymerase, and plasmids nucleotides, DNA template, DNA ligase, and plasmids nucleotides, DNA template, Taq polymerase, and primers restriction enzymes, DNA template, Taq polymerase, and primers nucleotides, DNA template, DNA ligase, and primersarrow_forwardLook 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_forward
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