Suppose you ordered a custom-synthesized PCR primer. The company that made the primer sent you a tube containing 50 nmol of dried primer – just a tiny bit of dry powder in the bottom of the tube. If you want to dissolve the primer in water to make a 100 μM primer solution, what volume of water should you add to the tube? (You can assume that the volume of water added is the final volume of the solution, because the DNA will take up almost no volume in solution.) (Show ALL your calculations on a piece of paper, take a picture and paste it in the space below).

Suppose you ordered a custom-synthesized PCR primer. The company that made the primer sent you a tube containing 50 nmol of dried primer – just a tiny bit of dry powder in the bottom of the tube. If you want to dissolve the primer in water to make a 100 μM primer solution, what volume of water should you add to the tube? (You can assume that the volume of water added is the final volume of the solution, because the DNA will take up almost no volume in solution.) (Show ALL your calculations on a piece of paper, take a picture and paste it in the space below).

Human Anatomy & Physiology (11th Edition)

11th Edition

ISBN:9780134580999

Author:Elaine N. Marieb, Katja N. Hoehn

Publisher:Elaine N. Marieb, Katja N. Hoehn

Chapter1: The Human Body: An Orientation

Section: Chapter Questions

Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...

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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.
If you are trying to set up a PCR with a total volume of 40uL and you have access to a stock solution of 5x buffer, how much 5x buffer will you be adding to the PCR? ___uL?
The photograph provided is of a 1 μg of 2-log ladder indicating the molecular size in kilobases (Kb) and amount of DNA per band in ng. Estimate the concentration of your PCR product (amplicon) in ng/μL by visual comparison. The expected size of your amplicon is about 600-bp (0.6 kb). ____μg/μL?
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.
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.
You are given a tube containing 275 ng of purified PCR product (DNA) that is 1262 bp long. How many picomoles of PCR product are in the tube? The average molecular weight of a deoxynucleotide monophosphate is 328 g/mol.
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: __________________________________
In a PCR-based crime scene investigation, similar to the one presented in the lab module with Brother Y and Brother X, there is a sample of DNA from a crime scene that is likely to belong to the guilty party. Based on the gel photo below, which shows the results of an electrophoresis gel following PCR amplification at one locus of 5 DNA samples - one crime scene sample and 4 suspects - which suspects can be excluded from this investigation? [Keep in mind that it is not possible for a heterozygous person to leave only one allele at a crime scene. If any one allele does not match, then that suspect is eliminated.] Choose all that apply.
You have added 4 μl of a 5 μM stock primer to a 25 μl PCR reaction. What is the amountof primer in nanomol that you have added to the reaction?
Why are DNA samples that are to be separated by gel electrophoresis always loaded at the cathode end of the power source? The sequencer in this lab experiment using a thin capillary tube to perform gel electrophoresis. Explain how this accomplishes the same task as traditional flatbed electrophoresis.
1) Prepare the following enzymatic reaction, present it in tabulated form. In a final volume of 30 ul, where buffer 4 (10 ml). How much volume of each reagent would be used and how much of water? Is there any problem? 2) The DNA pol 1 enzyme comes at a concentration of 50,000 U/ml. You have to prepare a 50 ug PCR reaction where you must use 0.05 U/ml reaction. You add 10 ul of PCR buffer, 2 ng of tempered DNA that is at a concentration of 0.5 ng/ul, primers (which are at 200 mM) so that each one remains at a concentration of 200 uM, Mg+2 that is 5 mM (10 X), enzyme and water. Present the table of all the reagents included in the reaction, the volumes of each one in ul. Present where the initial and final concentration of each reagent applies. Assume you have micropipettes for all values.
How many microliters of pGLO plasmid will you need for a PCR reaction is you need 200 nanograms and the plasmid concentration is 0.05 micrograms/microliter?