Below are several problems frequently faced by researchers when running the PCR. Give one(1) solution to each problem stated.1. No PCR product2. Multiple bands appeared after gel electrophoresis (you are amplifying just 1gene)3. Bright thick bands at the end of the agarose gel after electrophoresis4. Bands appeared at the negative control

The polymerase chain reaction or PCR is a method widely used in molecular biology. This method is…

Answered: Below are several problems frequently…

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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Similar questions

Below is the result of PCR products running on agarose gel electrophoresis. You expect to see one band, but the results show two bands, as shown below. Why is there a lower band? What are they? How can we avoid such a band?
A student is trying to add 9.0 pmol of primer mix to a 20.0 µL PCR. The primer mix is at a concentration of 50.0 µM, and the student determines that a serial dilution is required because directly adding the primer mix would require a volume less than 1.00 µL. The student takes 1.40 µL of the primer mix stock and mixes it with 25.0 µL of ultrapure H2O. Calculate the volume (in µL) of the intermediate solution required to add the correct amount of primer mix to the PCR.
In a typical PCR reaction what is happening in stages occurring at temperature ranges (a) 92–95°C, (b) 45–65°C, and (c) 65–75°C.
Please answer these two questions regarding PCR: a) Why do you need to perform PCR on DNA obtained from a crime scene? b) Why so forensic labs analyze non-coding DNA rather than genes?
You have a 100 micromolar stock solution of dNTPs in the freezer and are setting up a PCR reaction in which you need to have a final concentration of 20 micromolar dNTPs. In a reaction volume of 25 microliters, how much of your stock dNTPs do you add to each PCR reaction?
You have been performing a PCR reaction but your results aren't the greatest. Your Supervisor has told you that you should increase the concentration of Magnesium. What affect will this have on the reaction? a. The annealing temperature will not be affected but the enzyme activity will be affected. b. The annealing temperature will decrease. c. The denaturation temparture will have to be increased in the PCR protocol. d. The denaturation temparture will have to be decreased in the PCR protocol. e. The Annealing temperature will increase.
The best combination of regeants for a typical PCR reacion would include the following concentration of reagents: a. 1x buffer, 1.5 mMm MgCl2, 0.5nM primers, 10mMdNTPs, 2units Taq, 1010 copies of template b. none of these answers c. 1x buffer, 1.5 mMm MgCl2, 0.5pM primers, 10mMdNTPs, 2units Taq, 105 copies of template d. 1x buffer, 1.5 mMm NH3 Cl2, 0.5pM primers, 10mMdNTPs, 2units Taq, 102 copies of template e. 1x buffer, 1.5 mMm KCL2 , 0.5nM primers, 10mMdNTPs, 2units Taq, 103copies of template thank you!!
Imagine that you are producing a PCR product of approximately 2.1 kb in size with constant non-specific products which are two very light bands present above the desired product and another intense band below the desired product. Now for an experiment, you need very pure products out of the PCR process. To avoid non-specific product production, you want to take a different PCR approach. There are two PCR processes that your supervisor has recommended you which are: Touchdown PCR technique and Gradient PCR technique. Considering that you need the pure product only once for your future experiment, mention in no more than 5 sentences which PCR technique will you choose and why?
After you did the amplification by PCR, you run the product on 2% gel. Tell me how did you prepared the gel if the tray is 150 ml? how many milliliters of the TBE and how many grams of the Agarose? Draw a picture (On a paper or on computer) for the gel that you are expecting, the gene that you amplified is approximately 200 bp
What are the reasons why there needs to be more than 10 cycles in the PCR process? Name 3 reasons.
If you wanted to prepare only one PCR reaction, how much of each reagent would you add to the PCR tube?] What is the final volume of the individual PCR reactions we are making?
Ben used the following primer pairs to amplify the COI region of his DNA template: Afterwards, he used Agarose Gel Electrophoresis to visualize the PCR products. Based on the gel: a) Was he able to successfully amplify the desired region in all replicates? b) Are there errors in Ben's gel? Does he need to repeat his PCR?

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