14. Why denaturation stage is important in PCR? To answer the question, pleaseexplain:3.11. Recombinant DNA Technology189I) what is the basic principle of PCR?2) what are the components of PCR?3) what are the main stages of PCR?

The laboratory method that is used for the formation of multiple copies of DNA is quick succession…

Answered: 14. Why denaturation stage is important…

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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The idea behind PCR-based diagnostics is that a very small number of microbial genomes in a patient sample can be multiplied by PCR and more easily detected by the clinical team managing the patient’s care. Also, genetic-based diagnostics are very useful for viral infections because we don’t have biochemical tests, etc. to distinguish one virus from another (remember, viruses are metabolically inactive). However, a lot of work goes into the development of these tests. For instance, PCR requires primers that are complementary to the viral genome that is being copied. If primers are complementary to the target genome, what must scientists know to design primers that bind to the viral genome to be copied? (I mean this to be a general question; don’t look up the details of designing primers)
Options: I think it is 600 but I am not sure a) 1200 b) 800 c) 600 d) 500 e) 400
Q11) Explain, using the pCR 2.1 vector, what we would see if we try to grow cells on a plate with ampicillin and Xgal in each of these scenarios:a) transformation not successful (meaning the plasmid did not go inside the bacteria)b) transformation was successful, but no insert in LacZ gene (plasmid went in the bacteria but there was no inserted DNA in the LacZ gene of the plasmid)c) transformation and the ligation reactions were successful (got the plasmid in, and the plasmid is carrying extra DNA in the LacZ region, yay)
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
Put 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" the
1. Why is each of the following reagents required to be in the PCR reaction tube? Primers, Taq DNA polymerase, nucleotide mix, reaction buffer 2. Imagine that you forgot to add nucleotides to your PCR master mix. How would this affect the outcome of your PCR reactions? a) If you made the mistake described in the previous question, which control would give you unexpected results? (In other words, if you forgot to add nucleotides, which control reaction tube would have results that are different that they would be if you had prepared them correctly?) 3. If you found that there was DNA amplification in your negative control tube, what could be an explanation for that result?
What is the difference between PCR and real-time PCR, and how do you explain it?
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)
1.) What are the different temperatures used in PCR (polymerase chain reaction)? What happens at each temperature? 2.) What ingredients are used in PCR? What role does each ingredient have in replicating DNA? 3.) Why is the contamination of foreign DNA a particularly important problem when you do PCR?

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14. Why denaturation stage is important in PCR? To answer the question, please explain: 11. Recombinant DNA Technology 189 I) what is the basic principle of PCR? 2) what are the components of PCR? 3) what are the main stages of PCR?