You perform two single digests on the plasmid below, named pPER, with the restriction enzymes EcoRI and BamHI. One fragment, 6,000 base pairs in size, will result if the plasmid is digested with BamHI, which is illustrated in lane 2. The marker is in lane 4 and the sizes of markers are to the right of the gel in base pairs (bp). EcoRI, 5000- 0, 6000 PPER wwwwwwww 6000bp BamHI, 3000 EcoRI, 1000 Use the figures to answer the following questions. Question 3 1 EcoRI BamHI 2 4 ABC - -10,000 bp - 8,000 -6,000 -5,000 - 4,000 -3,000 D - 2,000 1,500 E 1,000 500 For the EcoRI single digest, determine the number and size of the resulting fragments by choosing the correct locations (letters) on the gel above.

You perform two single digests on the plasmid below, named pPER, with the restriction enzymes EcoRI and BamHI. One fragment, 6,000 base pairs in size, will result if the plasmid is digested with BamHI, which is illustrated in lane 2. The marker is in lane 4 and the sizes of markers are to the right of the gel in base pairs (bp). EcoRI, 5000- 0, 6000 PPER wwwwwwww 6000bp BamHI, 3000 EcoRI, 1000 Use the figures to answer the following questions. Question 3 1 EcoRI BamHI 2 4 ABC - -10,000 bp - 8,000 -6,000 -5,000 - 4,000 -3,000 D - 2,000 1,500 E 1,000 500 For the EcoRI single digest, determine the number and size of the resulting fragments by choosing the correct locations (letters) on the gel above.

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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You are studying a new plasmid, and you digest the plasmid with three restriction enzymes: Eco RI (E), Hindlll (H), and Xbal (X). You digest the plasmid DNA with each of the following combinations of enzymes and observe the results on an agarose gel. You are provided a partial plasmid map as shown below to the right. +18 a. What is the size of this plasmid in base pairs? b. What is the distance in base pairs between E1 and H? c. What is the distance in base pairs between E1 and X? d. What is the distance in base pairs between E2 and H? e. What is the distance in base pairs between E2 and X?
You are studying a new plasmid, and you digest the plasmid with three restriction enzymes: Eco RI (E), HindlII (H), and Xbal (X). You digest the plasmid DNA with each of the following combinations of enzymes and observe the results on an agarose gel. You are provided a partial plasmid map as shown below to the right. E+H E+X H+x Kb +4.3 +2.8 -+2.5 -2.0 - -1.8 +1.5 -1.0 12 F0.8 +0.5 a. What is the size of this plasmid in base pairs? b. What is the distance in base pairs between E1 and H? c. What is the distance in base pairs between E1 and X? d. What is the distance in base pairs between E2 and H? e. What is the distance in base pairs between E2 and X?
The following plasmid is digested with EcoR1 and Ndel restriction endonucleases. Agarose electrophoresis is then used for analysis. How many bands do you expect to see on the agarose gel after complete digestion? oic 6000 8000 bp 2000 EcoR1 EcoR1 i Bank Answer: Explanation: Ndel 4000
A plasmid is cleaved by restriction endonucleases and analyzed by agarose gel electrophoresis. Assume there is no supercoiling. Answer the following three questions about the plasmid. a. What is the size of plasmid? Explain briefly. b. How many HindIII sites are there in the plasmid? c. Is it possible to determine that how many EcoRI cut sites available in plasmid? Why? 1 2 3 4 Lane 1: DNA ladder (100 bp increments beginning at 100 bp), Lane 2: EcoRV, Lane 3: Smal, Lane 4: HindIII
Draw the gel electrophoresis results for the following 16 kb plasmid when the plasmid is cut withrestriction enzyme A, with restriction enzyme B and with both A and B restriction enzymes. The Aenzyme cuts at positions 0 and 9.2 kb and the sites are shown as white. (0 is at the top of the circle.) TheB enzyme cuts at positions 3.8, 7 and 13.2 kb and the sites are shown as gray. Show the electricalpolarity of the gel, the bands, and their sizes in your drawing. Make sizes consistent between gel lanes.Explain why the concept of charge density is important for gel electrophoresis.
Assume that a plasmid is 4700 base pairs in length and has restriction sites for a given restriction enzyme at the following locations: 800, 1400, 2900, and 3600. List the fragments by size that are ! expected when the plasmid is fully digested the restriction enzyme.
A plasmid DNA and a linear DNA (both of the same size) have one site for a restriction endonuclease. When cut and separated on agarose gel electrophoresis, plasmid shows one DNA band while linear DNA shows two fragments. Explain.
A result of agarose gel electrophoresis of plasmid DNA is shown below. The direction of the electrophoresis is indicated by the arrow head. Lane 1 shows the plasmid DNA with no restriction enzyme digestion. Lane 2 shows the plasmid DNA which has been digested by restriction enzyme ECORI. Given the fact that there are only relaxed and supercoiled forms existing in this plasmid, please annotate the bands in Lane 1 on this gel with their supercoiling status (supercoiled or relaxed) and explain why. Then based on the information from Lane 1 and Lane 2, analyze the number of EcoRI cleavage sites on this plasmid. Lane 1 2
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In the "Bacterial Transformation" experiment, why were there no fluorescence bacteria present in the other plates even though these were inserted by the +pGLO plasmid? (Note: Discuss the importance on the addition of ampicillin and arabinose to the medium of the genetically engineered bacteria). Limit your answer to 1-3 sentences only.
Assume that a circular plasmid is 3200 base pairs in length and has restriction sites for HindIII restriction enzyme at the following locations: 400, 700, 1400, 2600. Give the expected sizes of the restriction fragments following complete digestion.
you have transformed E.coli cells with a plasmid containing the gene for the enzyme beta-lactamase you have used 3µl of a stock solution of 2.5ng/µl DNA. You transformed 150/µl of competent cells, using the calcium chloride method. you have added 600µl of luria broth to the cells and then plated out 200µl on triplicate plates. upon counting the colonies, you got the following results: plate 1: 120 cfu plate 2: 35 cfu plate 3: 95 cfu please answer the following question: 1. what new characteristic will the cells have ater transformation? 2. how are you going to test for the new characteristic? 3. calculate the transformation efficiency of the experiment?