Biology: The Dynamic Science (MindTap Course List)
4th Edition
ISBN: 9781305389892
Author: Peter J. Russell, Paul E. Hertz, Beverly McMillan
Publisher: Cengage Learning
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Chapter 18, Problem 12TYK
Discuss Concepts A
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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.
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.
You 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).
Chapter 18 Solutions
Biology: The Dynamic Science (MindTap Course List)
Ch. 18.1 - What features do restriction enzymes have in...Ch. 18.1 - Prob. 2SBCh. 18.1 - What information and materials are needed to...Ch. 18.2 - What is a transgenic organism?Ch. 18.2 - Prob. 2SBCh. 18.3 - What is a restriction fragment length polymorphism...Ch. 18.3 - Prob. 2SBCh. 18.3 - Prob. 3SBCh. 18 - Prob. 1TYKCh. 18 - Prob. 2TYK
Ch. 18 - Why are antibiotic resistance markers such as ampR...Ch. 18 - After a polymerase chain reaction (PCR), agarose...Ch. 18 - A cDNA and a cloned fragment of genomic DNA share...Ch. 18 - Prob. 6TYKCh. 18 - Which of the following is not true of somatic cell...Ch. 18 - Prob. 8TYKCh. 18 - Prob. 9TYKCh. 18 - Prob. 10TYKCh. 18 - Prob. 11TYKCh. 18 - Discuss Concepts A forensic scientist obtained a...Ch. 18 - 13. Suppose a biotechnology company has developed...Ch. 18 - Prob. 14TYKCh. 18 - You learned in the chapter that an STR locus is a...
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- 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_forwardMatch the following terms with their definitions and label each component of the PCR mixture in the diagram (use the letters A-D):I. DNA polymeraseII. PrimersIII. NucleotidesIV. Genomic DNA template A. DNA that contains the target sequence that will be replicated using PCR.B. An enzyme that copies the DNA sequence.C. A mixture of 4 nucleotides (A,G,C, and T) that will be polymerized into the replicated DNA sequence.D. A short DNA sequence that allows the enzyme to bind and initiate polymerization.arrow_forwardPut the steps of one PCR cycle in the correct order: The PCR reaction mixture is heated to about 70 degrees, which is the optimum temperature for the polymerase to build the new strands of DNA, starting at the 3' end of the primer. The PCR reaction mixture is heated to 95 degrees Celsius, which denatures the double stranded template DNA. The PCR reaction mixture is cooled to about 50-55 degrees, which allows the primers to find their complementary site on the template and "anneal" thearrow_forward
- You 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_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_forwardplease solve this with step-by-step calculations and explanations.arrow_forward
- An analyst designed a pair of primers to detect the presence of Alu sequence in human DNA genome extract. The forward and reverse primers have the melting temperature at 70 and 50 Celsius degree respectively. Will the PCR work with the following PCR condition? Denaturing: 95 C for 1 min Annealing: 55 C for 1 min Extension: 72 C for 2 min Cycling: 35 cycles Yes. Annealing temperature at 55 is a standard in PCR. Yes. Difference in meting temperature of primer pair does not affect PCR. O No. There is chance that one of the primer won't anneal to the correct sequence. O No. DNA polymerase does not work at 72 Celsius degree.arrow_forwardThis is what the LAB FLOW would look like: CULTURE BACTERIA EXTRACT RNA RANDOM HEXAMER REVERSE TRANSCRIPTION OF RNA PCR OF lacZ GENE ELECTROPHORESIS If you wanted to prepare only one PCR reaction, how much of each reagent would you add to the PCR tube? 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_forwardA student carries out PCR using the following steps: step 1: 94C for 1 minuteStep 2: 60C for 30seconds Step 3: 72C for 30 seconds The student is examining a 1000-bp DNA fragment as part of their research project. If the limit of detection of this molecule 9 x 108 molecules, what is the minimum number of PCR cycles you would have to run to detect PCR product generated from a single molecule of the template? 2. Two double-stranded fragments of DNA are exactly the same length. At 89 C, fragment A has completely denatured which means the two strands have separated. At that temperature, fragment B is still double-stranded. How might these fragments differ to result in different denaturation temperatures?arrow_forward
- In a PCR reaction to amply a particular gene, 120 copy of the gene were initially used as DNA templates (this is the Nō). Theoretically, how many copies of this particular gene can be obtained after 25 PCR cycles? 4.03 × 10⁹ 2.01 × 10⁹ 1.02 × 10⁹ O 6.04 × 10⁹arrow_forwardYou are using a spectrophotometer to determine the concentration of DNA in a PCR product, but your samples need to be diluted to a 1:50 concentration in order to get an accurate reading. Which of the following dilutions would result in a 1:50 concentration? O 10uL of concentrated DNA into 490uL of DI water O 50uL of concentrated DNA into 950uL of DI water 1uL of concentrated DNA into 50uL of DI water O 5uL of concentrated DNA into 95uL of DI waterarrow_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
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