400 bp Hae III 380 bp Bam HI Eco RI 150 bp Hind II Plasmid DNA 2000 bp 250 bp 500 bp Eco RI 320 bp Eco RV a. Look up the restriction enzyme and indicate whether the restriction enzyme will produce a blunt or staggered cut. (1 mark) Note: Type either Blunt or Staggered Restriction enzyme Type of cut (Blunt vs Staggered) Eco RI b. Based on the plasmid map, list the size of fragments produced when the following restriction enzymes are used. Note: to answer correctly you must order the fragments from smallest to largest and separate each with a comma. There should be no spaces between numbers and commas. e.g. 400,500,530 Restriction Enzyme Eco RI and Hind II Eco RV and Bam HI Hae III, Eco RI and Bam HI Fragment size

400 bp Hae III 380 bp Bam HI Eco RI 150 bp Hind II Plasmid DNA 2000 bp 250 bp 500 bp Eco RI 320 bp Eco RV a. Look up the restriction enzyme and indicate whether the restriction enzyme will produce a blunt or staggered cut. (1 mark) Note: Type either Blunt or Staggered Restriction enzyme Type of cut (Blunt vs Staggered) Eco RI b. Based on the plasmid map, list the size of fragments produced when the following restriction enzymes are used. Note: to answer correctly you must order the fragments from smallest to largest and separate each with a comma. There should be no spaces between numbers and commas. e.g. 400,500,530 Restriction Enzyme Eco RI and Hind II Eco RV and Bam HI Hae III, Eco RI and Bam HI Fragment size

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

The map of plasmid pUC19 is shown below. The restriction site coordinate is the position of the 5’base on the top strand of each site sequence. The restriction enzyme sites are in bold type if there is only one site in pUC19. Please list the fragments in order of size, largest to smallest, which will result from a complete digestion by the restriction enzyme PvuII. Please list the fragments in order of size, largest to smallest, which will result from a complete digestion by the restriction enzyme DrdI.
Use the gel to answer the following questions. You will be constructing a map of the plasmid, pDiddy. There are NO sites where multiple enzymes cut. How many base pairs long is the plasmid? 3kb 2kb 4kb 5kb
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
Below is a plasmid with restriction sites for Baml and EcoRI, Several restriction digests were done using these two enzymes either alone or in combination. Hint: Begin by determining the number and size of the fragments produced with each enzyme. "kb" stands for kilobases, or thousands of base pairs. Plasmid Gel lanes I | III IV V 6 Kb Bam HI Bam HI. 20 Kb PGEN101 (20 Kb) 2 Kb 11 Kb Bam HI 8 Kb 8 Kb 4 Kb 6 Kb Eco RI 3 Kb 21. Which lane shows a digest with BamHl only? a. I b.I c. II d. IV e. V 22. Which lane shows a digest with EcoRLonly? a. I 23. Which lane shows the fragments produced when the plasmid was incubated with both EcoRI and BamH1? a. I b.I c. II d. IV e. V b. I c. II d. IV e. V Base pairs
Kha Vu Danels Include: 8DX : Safehy Jor bromie tnto lab repart! Name Section date sheet MAPPING PRACTICE #1 Below is a restriction map for the plasmid PGEN 101 (total length = 20 Kb). Using this map as a guide, give the number of restriction fraqments along with their associated lengths that would result from digesting PGEN 101 with the restriction enzymes EcoRI, BamHI and a combination of ECORI and BamHI. BamHI 3.2 Kb 1.7 Kb EcoRI BamHI PGEN 101 8.7 Kb 5.5 Kb .9 Kb EcoRI ECORI DIGESTION PERFORMED SIZES OF FRAGMENTS OBTAINED 10.4 kb , 0.9kb, 8.7 Kb EcoRI 3.2 Kb, 16. 8kb BamHI EcoRI + BamHI
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.
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+H E+X H+X Kb +4.3 +2.8 +2.5 - +2.0 -- -1.8 -1.5 12 +1.0 +0.8 +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 figure attached represents the structure of a pUC19 plasmid. A 1000 bp fragment was inserted into a Sma I-cut plasmid. The Sma I restriction enzyme site is located in the MCS region of the plasmid. What does the MCS name stand for?
U have the plasmid pUC18/19, which is a circular plasmid that consists of 2686 bp. What would the number of and length of the fragments be if you cut the plasmid with the following restriction enzymes or combination of enzymes? Give a schematic representation of the digestions. PscI & GsuI ______________________________________________________________ ScaI, PdmI & BsaXI ______________________________________________________________ ScaI, SspI & EheI ______________________________________________________________
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.
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
How many fragments and how big each fragment should be after digesting the plasmid with BamHI? How many fragments and how big each fragment should be after digesting the plasmid with PvuI? How many fragments and how big each fragment should be after digesting with both enzymes? Show your calculation/explanation.