What would be the final primer concentration if 0.5 ul of 10 uM primers were added toa PCR reaction with a final volume of 20 ul?O0.25 uMO 20 LMO 0.2 uMO s uM

Introduction: A primer is a short strand of DNA or RNA that is usually about 18-22 bases and serves…

Answered: What would be the final primer…

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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Our PCR samples already contain loading dye, but sometimes this isn’t the case. If your samples didn’t already contain dye and you wanted to load your PCR sample onto an agarose gel, you’d need to add loading dye to the proper concentration. There is a 6X loading dye available for use; how many µl of this loading dye will you add to 10 µl of your sample so that it is at a 1X working concentration? Show your work.
What is the sequence of the sample DNA submitted for sequencing given the gel electrophoresis profile from the Sanger sequencing method? ddATP ddCTP ddGTP ddTTP
You have run a gel with 5 uL of amplified DNA in addition to 1 uL of 6X loading dye. Comparing the brightness of this band to your marker, you estimated that your amplified DNA is approximately 62 ng. What was the concentration of your amplified DNA that was loaded? You do not need to account for the volume of loading dye in your calculations.
Draw your own PCR diagram to show the role of each component and relevance of each temperature shift during the PCR reaction. What’s happening at each of those temperatures and why is it important it changes?
The sequence 5' GCCTAATGCGTTCATAATGGCGTTTGCCACGGACGTAAAGTCGT 3' represents 50% of a PCR product which is cloned into a 1.5 kb plasmid for bacterial expression. What would the agarose gel look like, including a lane for markers, and where would the following pieces of DNA run: 1. the PCR insert 2. the empty uncut plasmid 3. the empty plasmid cut with a single restriction enzyme 4. the cloned plasmid containing the insert
Using the designed forward and reverse primer from Question 23-30, you performed PCR to amplify the CO1 gene of S. tawilis. Then, you verified the PCR product using the Agarose Gel Electrophoresis. Here is the result of the AGE. M-Marker L1-PCR Product ML1 You noticed that there are many bands in your PCR product. What does this indicate? Choose the best answer. The primers were not specific to CO1 gene. The annealing temperature is too high. The templated DNA is not enough. There is a formation of primer dimers. The primer length is too short. The melting temperature is too high.
Run a barcoding protocol with rbcL gene-specific primers. Use 30 cycles for your PCR amplification profile. Complete the table below for your PCR cocktail.
Copy and paste the link below and watch the video on Youtube and Answer the Questionshttps://www.youtube.com/watch?v=g-dNJdOvBM4 Polymerase Chain Reaction Questions: 1. What are the materials used for the polymerase chain reaction? 2. Draw a schematic diagram of the procedure in PCR. 3. Why is it important to design the primers at the start of the laboratory procedure? 4. What are the components of the PCR buffer and what is its pH range? What is the purpose of the buffer? 5. What is the use for magnesium chloride? 6. How much template DNA is added? What is the concentration of the primers? 7. At what temperatures does denaturation, annealing and extension occur? Why are the processes placed in that temperature? 8. In this particular PCR experiment, how many cycles was used? 9. Can this PCR be used on its own to find out if a person has Covid or not on its own? Why or why not?
please solve this with step-by-step calculations and explanations.
All things considered, the most important factor with respect to successful PCR is Multiple Choice primer design annealing temperature extension time quantity of template
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?
What are the reasons why there needs to be more than 10 cycles in the PCR process? Name 2 reasons.

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What would be the final primer concentration if 0.5 ul of 10 uM primers were ad a PCR reaction with a final volume of 20 ul? O 0.25 uM O 20 uM 0.2 uM OS uM 1 M