**PCR Result Analysis****Scenario:**You are performing PCR (Polymerase Chain Reaction) for the first time using new primers that are 25 nucleotides in length, with an estimated melting temperature of 65°C, to amplify a 200 nucleotide gene. After conducting PCR, you run an agarose gel on the samples and observe a faint band at 25 nucleotides.**Question:**What is the best possible explanation for the observed results?**Answer:**- **The primers hybridized to multiple sites on the DNA template.** (Highlighted as the correct explanation) Additional Explanations (not selected):- Too much template DNA was added to the PCR mixture.- The primers dimerized, preventing DNA transcription from occurring.- A nuclease was in the solution, causing degradation of the DNA.**Diagram Explanation:**There is no diagram or graph in this text section to explain. The primary focus is on the interpretation of the PCR result as evident from the band pattern in the gel electrophoresis. The faint band at 25 nucleotides suggests that primers are binding to unintended sites, causing amplification of shorter, non-target fragments.

PCR is the process by which we increase the number of a specific nucleotide sequence. The PCR…

Answered: You are performing PCR for the first…

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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An 9 kb circular plasmid is cut with the EcoRI restriction enzyme, and the reaction products are run on a DNA gel and stained with ethidium bromide. Bands of 1, 3 and 5 kb are seen. How many EcoRI sites are in the plasmid? Choose the one answer that is most correct. a) At least 2 b) At least 3 c) At least 1 d) None e) At least 4 f) At least 5
Options: I think it is 600 but I am not sure a) 1200 b) 800 c) 600 d) 500 e) 400
The sequence of a template DNA is GGGCCATTCGAACGTCCGAAAATGCCCCTGAATGAAAATTTTGGCCC. The sequence of a primer used for PCR amplification of the above DNA is CCCGGTAAGCTT. Where is the 5' end for the template and primer, respectively? answer options: 1) Left, left 2) Right, left 3) Left, right 4) Right, right 5) Right, left
Recently, I am designing primers specific for Lactobacillus sp. (partial sequence of the 16s rRNA gene) and can get successful amplification on my conventional PCR. Then I am going to quantify by RT-PCR using SyBr Green Reagent in Step One Plus Instrument. By using the same run condition as the conventional PCR, I face the problem that many of my samples got a bad melting curve. Anyone have the same experience as me or suggestion please let me know. Thanks in advance.
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
We usually think of enzymes as being most active at around 37°C, yet in PCR the DNA polymerase is subjected to multiple exposures of relatively high temperatures and seems to function appropriately at 65–75°C. What is special about the DNA polymerase typically used in PCR?
PCR is an exponential copying of the template strands and can be represented by the function: y = a * 2n, where a is the initial number of template copies and n is equal to the number of cycles PCR has gone through. How many DNA fragments would be produced after: 15 cycles? 13 cycles with 13 starting template strands? 29 cycles with 32 starting template strands?
A more modern molecular technique to RFLP fingerprinting is called Amplified Fragment Length Polymorphisms (AFLPs). In AFLP analysis, restriction enzymes are again used to digest genomic DNA into multiple fragments. Next, adapters complementary to restriction site overhangs are ligated to the fragments using an enzyme called DNA ligase. These adapters are complementary to primers used to amplify the fragments using the polymerase chain reaction (PCR). Can you think of any potential benefits of AFLP analysis over RFLP? Explain your reasoning.
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?
PCR is a molecular biology technique where template DNA is amplified using a primer and oligonucleotides. The reaction is catalyzed by a thermostable DNA polymerase and in a particular reaction, the template strands are denatured at 95˚C. For strand hybridization, the melting temperature is 55˚C. What do you predict about the average duration of H bonds at the high temperature in comparison to the low temperature?

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