4. Create a PCR master mix for the following: 48 samples, 25 uL total volume per reaction, 2 uL template DNA volume, 2.5mM MgCl, 0.5 uM each primer. Your stock concentrations are: reagent mix (2X, does not contain MgCl,), primers (F/R; 10 µM), MgCl, (50 mM). The master mix calculation must include a 10% overage, and all answers are to be given to one decimal place.

4. Create a PCR master mix for the following: 48 samples, 25 uL total volume per reaction, 2 uL template DNA volume, 2.5mM MgCl, 0.5 uM each primer. Your stock concentrations are: reagent mix (2X, does not contain MgCl,), primers (F/R; 10 µM), MgCl, (50 mM). The master mix calculation must include a 10% overage, and all answers are to be given to one decimal place.

Biology: The Unity and Diversity of Life (MindTap Course List)

15th Edition

ISBN:9781337408332

Author:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr

Publisher:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr

Chapter15: Studying And Manipulating Genomes

Section: Chapter Questions

Problem 16SQ: Match the method with the appropriate enzyme. _____ PCR a. Taq polymerase _____ cutting DNA b. DNA...

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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.
1) Which technique is best suited to determining which genes are activated in a bacterium during infection while causing disease in a person. a) SDS-page b) microarray analysis c) RFLP analysis d) clone library analysis 2)Which of the following is not an application of PCR? a) Determine if two people are related. b) Identify a bacterial pathogen in a patient sample. c) Determine the gene sequence of the gene that codes for a bacterial enterotoxin. d) These are all applications of PCR.
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1. a) A technologist has a 20µl sample of DNA and adds 5µl of loading dye before adding the total volume into an agarose gel. What was the original concentration of the loading dye? a. 4x b. 5x c. 6x d. 7x e. None of the above 1. b)A technologist adds 6uL of 5x loading dye to a DNA sample before adding the total volume into the agarose gel. How much DNA sample was combined with the loading dye? 12uL 16uL 20uL 24uL 28uL
1. The final amount of each primer required in a PCR reaction is 0.025 nanomol. If the total volume of the PCR reaction is equal to 100 µl and the stock concentration of each primer is equal to 10 µM. Calculate the volume of stock primer that needs to be added in order to ensure a primer amount of 0.025 nanomol. 2. A 250 ml phosphate buffer solution contains 204.12 µg KH2PO4 and 87.09 µg K2HPO4, respectively. Determine the molarities in micromolar (µM) of the abovementioned compounds in the phosphate buffer solution. [Mw KH2PO4 = 136.08 g/mol and Mw K2HPO4 = 174.18 g/mol] 3. You are required to use the pET 28b expression vector in a cloning experiment. If the vector is 5.368 kilobasepairs in length and the mass of the DNA that you have added represent 8.5 x 109 molecules. Determine the mass of vector DNA that you have added to your reaction? [1basepair ~ 660 Da or g/mol, Avogadro’s constant = 6.023 x 1023 molecules/mol]
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Your amplicon from PCR was subjected to AGE for analysis. You are shown the image of the gel loaded with the following lanes: (A) negative control, (B) size ladder (1, 2, 3, 4, 6, and 10 kb), (C) gDNA extract, (D) PCR amplicon. However, due to mishaps while loading the gel with the samples, you are not sure which lane is which. You are shown a diagram of the obtained gel below. a. Label each lane of the gel. Write only the corresponding letters in the wells above. b. Above each band in the size ladder, write its size (in kb). c. Approximate the size (in kb) of the polystyrenase gene. Write your answer above the band corresponding to the gene. Bonus: If you wish to identify the bacterial species in this scenario, what gene is most commonly and routinely sequenced? Answer: __________________________________
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Please answer these two questions regarding PCR a) First question you must use the image: What is the DNA profile for suspect B for the TH01 locus? b) What determines the target sequence amplified in PCR?
6. Which of the following steps are catalyzed by Taq DNA polymerase in a PCR reaction?a. Denaturation of template DNAb. Annealing of primers to template DNAc. Extension of primer end on the template DNAd. All of the above Please also explain the answer properly and refer to NCERT XII (chapter 11) textbook.