What would be the effect of performing a RT-PCR with the following ingredients: an mRNA template,appropriate primers, dNTPs, heat-stable reverse transcriptase and human DNA polymerase?Select one:a. The PCR would occur, but with a high mutation rateb. The PCR reaction will not commencec. Non-specific PCR of random templates will occurd. The RNA template would be converted to DNA, but the DNA segment would not be amplifiedPCR would proceed normallyе.

The ingredients required for RT PCR are given below Reverse transcriptase enzyme mRNA template.…

Answered: What would be the effect of performing…

Biochemistry

9th Edition

ISBN:9781319114671

Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Chapter1: Biochemistry: An Evolving Science

Section: Chapter Questions

Problem 1P

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Match 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.
If Susie collects her cheek cells, but accidentally dumps the cell pellet into the waste container in step 4 of the DNA collection procedure, and doesn't realize it, what is the problem with her sample, moving forward, if any? A Susie has no cells in her sample, so no DNA can be prepared from her sample. The PCR reaction will not contain template DNA. B No problem. Susie can expect the PCR reaction to work fine.
What would be the expected effect on the PCR reaction, if you increased the temperature of the annealing phase and the length of the elongation phase? O Accuracy will be reduced, but yield will be increased Accuracy and yield will be increased OAccuracy will be increased, but yield will be decreased Accuracy and yield will be reduced
Primer annealing is an important aspect of PCR. The annealing step of the cycle usually takes place at around 45-55°C and lasts for only 20-30 seconds. Which of the following statements best explains why the annealing time has to be so brief? OA. A longer annealing time would prevent primers binding to specific sites. OB. The short time frame prevents primers from binding to each other. OC. If the time were any longer, DNA polymerase would begin to denature. OD. The short time frame minimizes complementary template strands base pairing with each other. Reset Selection
Which of the following statements about PCR or RT-gPCR is correct? 1. Fluorescence is detected when the TagMan probe is cleaved by Taq polymerase (AMPLITAQ). 2. PCR uses the natural process of transcription to amplify DNA. 3. Both PCR and RT-qPCR require forward and reverse primers. 4. Fluorescence is detected when the reporter dye is in close proximity to the quencher dye. A. 1, 2 and 3 B. 1 and 3 C. 2 and 4 D. 4 only E. All of 1, 2, 3 and 4 are correct
What are the advantages of qPCR (RT-PCR) compared to conventional PCR? Choose all that apply a. human error is reduced as there are fewer human interactions with the samples b. you can visualize the results as the process is running c. samples can be compared as to the amount of template DNA in the original sample d. more samples can be run in a day by one person
During a PCR reaction, all the following events occur except: A. Hybridization B. Binding of a 3’OH group in the active site of DNA polymerase C. Addition of nucleotide monomers on the 3’ end of the template DNA D. DNA denaturation E. Semi conservative replication
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 600
Which of the following reagents is NOT required for a PCR? a. DNA template b. DNA Polymerase c. All four dNTPs d. small amount of ddNTPs e. Pair of primers
What would be the outcome if the primers used in a polymerase chain reaction have lower GC content (<40 %), shorter, and more variable than the intended oligonucleotide sequence? a.The PCR reaction will cease after the first cycle b.The reaction will yield a mixture of non-specific products. c.All of these d.The PCR reaction will not start. e.The reaction will yield a single short PCR product.
In a reaction tube, a PCR reaction recreates the DNA replication process inside a cell. Therefore, a PCR reaction needs to have the following essential components: DNA polymerase DNA template Forward and Reverse PCR primers dNTPs, including dATP, dCTP, dTTP, and dGTP MgCl2 True O False
Choose the one answer that fits best. Which statement regarding PCR is NOT correct (videos)? a. PCR requires a copy of RNA that serves as a template b. Taq polymerase adds nucleotides to the primers and creates a complementary strand of DNA c. Annealing requires cooler temperatures than denaturation d. Repeated cycles of denaturation, annealing and extending DNA strands creates many identical copies of DNA e. PCR is a quick way of using minute quantities of DNA to create millions of copies

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