Homework Answers
As the DNA is linear & after digestion with PstI & SmaI produces 2 & 3 restriction fragments respectively, PstI & SmaI will have 1 & 2 restriction cut sites respectively.
At first, we draw restriction map based on only PstI digestion. This is shown in below picture.
Next, we consider PstI+SmaI digestion. We find that 2 kb & 8 kb fragments of only SmaI digestion remain intact in PstI+SmaI digestion. Further, 9 kb fragment of only SmaI digestion divided into two fragments of 4 kb & 5kb. So, PstI restriction cut site must be present in the 9 kb fragment of only SmaI digestion. Based on this, the final map is drawn below.
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A linear fragment of DNA is cleaved with the individual restriction enzymes Pst and Smal, and...
A linear fragment of DNA is cleaved with the individual restriction enzymes Hindill and Smal, and...
A linear fragment of DNA is cleaved with the individual restriction enzymes Hindill and Smal, and then with a combination of the two enzymes. The fragments obtained are: Eco RV 2 kb, 3.5 kb, 9.5 kb Bam HI 6 kb, 9 kb Eco RV and Bam H12 kb, 3.5 kb, 4 kb. 5.5 kb Draw a restriction map of the DNA fragment. (5 marks). Model of example restriction map to show you how to give your answer: Eco R1 Hind...
A DNA molecule is cleaved with Hind III, Pst II, or the two enzymes together. The...
A DNA molecule is cleaved with Hind III, Pst II, or the two enzymes together. The accompanying diagram shows the resulting electrophoresis gel, with band sizes indicated. Is the DNA molecule in question linear or circular? Linear circular Using the information in question 6. determine where the Pst 1 restriction site is located. Pstl cleave within the 7kb HindM fragment or within the 3kb HindM fragment'; 7kb fragment 3kb fragment neither fragment both fragments
You digest a 10Kb Linear ECoRI dna fragment with TWO restriction enzymes and obtain the following...
You digest a 10Kb Linear ECoRI dna fragment with TWO restriction enzymes and obtain the following data Hind iii..... 3kb, 7kb BamHi ... 2 kb and 8kb HInd iii + bamHI ..... 1kb, 2kb, 7kb Draw a restriction map of this DNA fragment, labeling the sites for EcoRI, bahHI, and HIndiii
Restriction Mapping of a Linear DNA • 2 DNA is a linear and double-stranded DNA isolated...
Restriction Mapping of a Linear DNA • 2 DNA is a linear and double-stranded DNA isolated from bacteriophage lambda. This bacteriophage is a virus that infects E. coli and can undergo specialized transduction (see the transduction lab lecture). Assuming you obtained the following fragments from the restriction digestion. Draw the restriction map for the EcoRI and Hindi II enzymes in the 2. DNA. Fragment Size EcoRI 15300 6200 5250 2100 EcoRI + HindIII 15300 4400 4000 2100 1800 Hind!!! 17100...
5. You have a linear DNA fragment and you wish to generate a restriction map using...
5. You have a linear DNA fragment and you wish to generate a restriction map using Mstl and EcoR1. When the framgent is digested with Mst1 and the DNA is run on a gel you observe a 4kb and a 6kb fragment. When the DNA is digested with EcoR1, 1kb, 4kb and 5kb fragments are generated, A double digest using both enzymes produce 1kb, 2kb, 3kb and 4kb fragments. Explain these results and draw a restirction map consistent with these...
Restriction Mapping Below is a restriction map for the plasmid PGEN101 (total length - 20 kb)....
Restriction Mapping Below is a restriction map for the plasmid PGEN101 (total length - 20 kb). Using this map as a guide, give the number of restriction fragments along with their associated lengths that would result from digesting PGEN101 with the restriction enzymes EcoRI, BamHII, anda combination of EcoRI + BamHI. BamHI BamHI BamHI / PGEN101 (20 kb) Mb EcoRI Digest Performed Size Emments Obtained EcoRI........ BamHI.. EcoRI + BamHI.... Two freshmen college students, interested in becoming gene jocks, performed...
A linear DNA molecule that is 3250 bp long is digested with restriction enzyme A alone,...
A linear DNA molecule that is 3250 bp long is digested with restriction enzyme A alone, restriction enzyme B alone, or a combination of enzymes A and B to produce the following fragment sizes. Construct a restriction map of the DNA fragment. How many times does enzyme A cut the linear DNA molecule? If you are unable to place both enzymes A and B on the map, show one possible arrangement of enzyme A cognition sites on the DNA molecule.
The figure below shows a restriction map of a segment of a DNA molecule. Eco refers...
The figure below shows a restriction map of a segment of a DNA molecule. Eco refers to locations where the restriction endonuclease EcoRI cuts the DNA, and Pst refers to locations where the restriction enzyme Pst cuts the DNA. Potential restriction sites are numbered 1-6. Distances between restriction sites are shown on the bottom scale in base pairs (bp). The thick line represents the part of the molecule that has homology with a probe. Eco Pst Eco Pst Eco Pst...
1) Imagine that you had two restriction enzymes and a known segment of linear DNA 160...
1) Imagine that you had two restriction enzymes and a known segment of linear DNA 160 kb long: Enzyme A cuts at location(s): 20 kb, 45 kb 70 kb and 110 kb. Enzyme B cuts at location(s): 15 kb and 140 kb. Based on this information, first construct a linear map of this DNA showing the positions of these restriction enzyme cut sites. In the space below.draw what the gel would look like given the following lanes: (8 points total)...
on the me but not with the larger fragments on o t and more and record...
on the me but not with the larger fragments on o t and more and record med in the h 3. Determine the distracted for each bund data bere 4. Determine the distance traveled for each and generated in the double digest and record data bere tion Endonuclease Digestion and Gel Electrophoresis of DNA 185 VI. RESTRICTION MAPPING The different DNA fragments generated by different map DNA. For example, specific DNA on 2000 nucleotides in las Rl or Hind and...
A linear fragment of DNA is cleaved with the individual restriction enzymes Hindill and Smal, and then with a combination of the two enzymes. The fragments obtained are: Eco RV 2 kb, 3.5 kb, 9.5 kb Bam HI 6 kb, 9 kb Eco RV and Bam H12 kb, 3.5 kb, 4 kb. 5.5 kb Draw a restriction map of the DNA fragment. (5 marks). Model of example restriction map to show you how to give your answer: Eco R1 Hind...
A DNA molecule is cleaved with Hind III, Pst II, or the two enzymes together. The accompanying diagram shows the resulting electrophoresis gel, with band sizes indicated. Is the DNA molecule in question linear or circular? Linear circular Using the information in question 6. determine where the Pst 1 restriction site is located. Pstl cleave within the 7kb HindM fragment or within the 3kb HindM fragment'; 7kb fragment 3kb fragment neither fragment both fragments
Restriction Mapping of a Linear DNA • 2 DNA is a linear and double-stranded DNA isolated from bacteriophage lambda. This bacteriophage is a virus that infects E. coli and can undergo specialized transduction (see the transduction lab lecture). Assuming you obtained the following fragments from the restriction digestion. Draw the restriction map for the EcoRI and Hindi II enzymes in the 2. DNA. Fragment Size EcoRI 15300 6200 5250 2100 EcoRI + HindIII 15300 4400 4000 2100 1800 Hind!!! 17100...
5. You have a linear DNA fragment and you wish to generate a restriction map using Mstl and EcoR1. When the framgent is digested with Mst1 and the DNA is run on a gel you observe a 4kb and a 6kb fragment. When the DNA is digested with EcoR1, 1kb, 4kb and 5kb fragments are generated, A double digest using both enzymes produce 1kb, 2kb, 3kb and 4kb fragments. Explain these results and draw a restirction map consistent with these...
Restriction Mapping Below is a restriction map for the plasmid PGEN101 (total length - 20 kb). Using this map as a guide, give the number of restriction fragments along with their associated lengths that would result from digesting PGEN101 with the restriction enzymes EcoRI, BamHII, anda combination of EcoRI + BamHI. BamHI BamHI BamHI / PGEN101 (20 kb) Mb EcoRI Digest Performed Size Emments Obtained EcoRI........ BamHI.. EcoRI + BamHI.... Two freshmen college students, interested in becoming gene jocks, performed...
A linear DNA molecule that is 3250 bp long is digested with restriction enzyme A alone, restriction enzyme B alone, or a combination of enzymes A and B to produce the following fragment sizes. Construct a restriction map of the DNA fragment. How many times does enzyme A cut the linear DNA molecule? If you are unable to place both enzymes A and B on the map, show one possible arrangement of enzyme A cognition sites on the DNA molecule.
The figure below shows a restriction map of a segment of a DNA molecule. Eco refers to locations where the restriction endonuclease EcoRI cuts the DNA, and Pst refers to locations where the restriction enzyme Pst cuts the DNA. Potential restriction sites are numbered 1-6. Distances between restriction sites are shown on the bottom scale in base pairs (bp). The thick line represents the part of the molecule that has homology with a probe. Eco Pst Eco Pst Eco Pst...
1) Imagine that you had two restriction enzymes and a known segment of linear DNA 160 kb long: Enzyme A cuts at location(s): 20 kb, 45 kb 70 kb and 110 kb. Enzyme B cuts at location(s): 15 kb and 140 kb. Based on this information, first construct a linear map of this DNA showing the positions of these restriction enzyme cut sites. In the space below.draw what the gel would look like given the following lanes: (8 points total)...
on the me but not with the larger fragments on o t and more and record med in the h 3. Determine the distracted for each bund data bere 4. Determine the distance traveled for each and generated in the double digest and record data bere tion Endonuclease Digestion and Gel Electrophoresis of DNA 185 VI. RESTRICTION MAPPING The different DNA fragments generated by different map DNA. For example, specific DNA on 2000 nucleotides in las Rl or Hind and...
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