An analyst designed a pair of primers to detect the presence of Alu sequence in human DNAgenome extract. The forward and reverse primers have the melting temperature at 70 and 50Celsius degree respectively. Will the PCR work with the following PCR condition?Denaturing: 95 C for 1 minAnnealing: 55 C for 1 minExtension: 72 C for 2 minCycling: 35 cyclesYes. 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.

PCR stands for a polymerase chain reaction. PCR is carried out under in-vitro or test tube…

Answered: An analyst designed a pair of primers…

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

A successful PCR experiment often depends on designing the correct primers. In particular, the T m for each primer should be approximately the same. What is the basis of this requirement?
The final amount of each primer required in a PCR reaction is 25 picomol. If the total volume of the PCR reaction is equal to 100 µl and the stock concentration of each primer is equal to 0.0025 mM. Calculate the volume of stock primer that needs to be added in order to ensure a primer amount of 25 picomol.
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
Ideal primers for the PCR reaction should have the following feature: they should have a high A-T content the forward primer should be palindromic to the reverse primer they should anneal rapidly only on one target in the DNA template they should have a high G-C content
To generate large amounts of DNA to manufacture gene therapy payloads or to be able to see them on gel electrophoresis, specific sequences can be amplified by PCR. For a sequence of a 100 base pairs, calculate the number cycles of PCR required to generate 1 ng of DNA, when starting from a single copy. Note that the molar mass of an average DNA bp is 600 g/mol.
The PCR primers are at a 10 uM stock concentration, how much of each primer would you add to get 0.05 uM final concentration of each in a final PCR reaction of 100 ul.
Explain why DNA ladders are usually included during gel electrophoresis. One aspect of PCR that can be modified is the annealing temperature. In general, higher annealing temperatures show more specificity towards a single template, whereas lower annealing temperatures show less specificity and may bind to multiple regions throughout the genome. Discuss how using an annealing temperature that is too high or too low might influence the results of a PCR assay (and gel electrophoresis results) such as the one used in this study.
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 process of PCR essentially revolves around three phases. Briefly describe these phases and the events that occur in them. Take note the temperature on which these phases take place.
Why is the company Qiagen has more refined DNA extraction steps than a normal Strawberry DNA extraction practical? Summary of Qiagen DNA extraction steps Add ATL buffer and grind with sample. Add 20 microliters of enzyme Proteinase K to degrade protein into a 1.5-2ml microcentrifuge tube. Add 200 microlitres AL lysis buffer, and mix by vortexing for 5–10 seconds, which breaks cell membrane allowing DNA to be released. Incubate the sample at 56 degrees for 10 minutes. Mix the cell lysate with 200 microlitres ethanol by pipetting it at the side of the microcentrifuge wall so DNA precipitates. The DNA forms a white layer and the remaining liquid is discarded. Pipet the mixture into DNeasy Mini spin column placed in a 2 ml collection tube. Centrifuge for a minute at 8000 rpm. Place the mini spin column into a 2 ml collection tube, add 500 µl Buffer AW1, and centrifuge for 1 min at 8000 rpm. Then add it to a new 2 ml collection tube (provided), add 500 µl Buffer AW1, and centrifuge for 1…
You want to set up 50 µl total volume of a PCR reaction. You have a microfuge tube of Forward primer (25 µM), and each PCR reaction requires a final of 1 uM of primer. You have a microfuge tube of Reverse primer (25 µM), and each PCR reaction requires a final of 1 µM of primer. How much of the primers would you add? Select all that apply 2 ul of Reverse primer 2 ul of Forward primer 5 ul of Forward primer 5 ul of Reverse primer
5' 3' O 60°C 5' O 95°C Primer 1 O 75°C ORF For your PCR reaction above, the primer set has a much higher AT (low GC) content than what would be considered normal. Due to this difference in AT content, the optimal annealing temperature for the primers is going to have to be altered from the "normal" temperature cycle: 95°C 60°C ⇒ 75°C Which of the temperatures will have to be altered to accommodate this difference the primer set? Primer 25¹ 3' 5' annealing with

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