Bsal 6. On the left is a diagram of the pUC19 plasmid, which is composed of 2,686 bp. Numbers on the plasmid and next to each restriction enzyme cut site represent a base pair (bp) measurement from an arbitrary starting point (0) going in the clockwise direction. On the right is a diagram of an electrophoresis gel. A standard molecular weight (MW) ladder is in the far-left lane, labeling the position of DNA fragments of known sizes from 0.5-10 kb. Lanes 1-5 are DNA bands observed from digestion of the plasmid with one or more of the restriction enzymes. 2500 11500 or PUC19 2686 bp Sspl (1266) P HindIII (632) BamHI (662) EcoRI (683) kb 10.0. 8.0 6.0 5.0. 4.0- 3.0 2.0 1.5 1.0 <0.5- MW 1 |||||| || | || 2 3 4 5 a. Which lane(s) shows the DNA fragmentation pattern produced by digestion of the plasmid with either EcoRI, Bsal, OR Ssp1? Briefly explain your answer. b. Cutting this plasmid with restriction enzymes Bsal AND Ssp1 would result in DNA fragments of what sizes? Which lane(s) of the gel shows this expected DNA fragmentation pattern? c. If you want to clone a piece of DNA into the MCS (labeled in blue), which restriction enzyme(s) could you use? d. After digestion of the plasmid and the DNA insert with this restriction enzyme, what enzymatic reaction is necessary to generate the desired recombinant plasmid DNA?
Bsal 6. On the left is a diagram of the pUC19 plasmid, which is composed of 2,686 bp. Numbers on the plasmid and next to each restriction enzyme cut site represent a base pair (bp) measurement from an arbitrary starting point (0) going in the clockwise direction. On the right is a diagram of an electrophoresis gel. A standard molecular weight (MW) ladder is in the far-left lane, labeling the position of DNA fragments of known sizes from 0.5-10 kb. Lanes 1-5 are DNA bands observed from digestion of the plasmid with one or more of the restriction enzymes. 2500 11500 or PUC19 2686 bp Sspl (1266) P HindIII (632) BamHI (662) EcoRI (683) kb 10.0. 8.0 6.0 5.0. 4.0- 3.0 2.0 1.5 1.0 <0.5- MW 1 |||||| || | || 2 3 4 5 a. Which lane(s) shows the DNA fragmentation pattern produced by digestion of the plasmid with either EcoRI, Bsal, OR Ssp1? Briefly explain your answer. b. Cutting this plasmid with restriction enzymes Bsal AND Ssp1 would result in DNA fragments of what sizes? Which lane(s) of the gel shows this expected DNA fragmentation pattern? c. If you want to clone a piece of DNA into the MCS (labeled in blue), which restriction enzyme(s) could you use? d. After digestion of the plasmid and the DNA insert with this restriction enzyme, what enzymatic reaction is necessary to generate the desired recombinant plasmid DNA?
Human Anatomy & Physiology (11th Edition)
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Question
Transcribed Image Text:2005) Bsal
6. On the left is a diagram of the pUC19 plasmid, which is composed of 2,686 bp. Numbers on the
plasmid and next to each restriction enzyme cut site represent a base pair (bp) measurement
from an arbitrary starting point (0) going in the clockwise direction.
On the right is a diagram of an electrophoresis gel. A standard molecular weight (MW) ladder is
in the far-left lane, labeling the position of DNA fragments of known sizes from 0.5-10 kb. Lanes
1-5 are DNA bands observed from
digestion of the plasmid with one
or more of the restriction enzymes.
2500
11500
ori
PUC19
2686 bp
Sspl (1266)
9
B
-HindIII (632)
BamHI (662)
EcoRI (683)
kb
10.0,
8.0
6.0.
5.0
4.0
3.0
2.0
1.5
1.0
<0.5
MW 1
| ||
2 3 4 5
a. Which lane(s) shows the DNA fragmentation pattern produced by digestion of the plasmid with either
EcoRI, Bsal, OR Ssp1? Briefly explain your answer.
b. Cutting this plasmid with restriction enzymes Bsal AND Ssp1 would result in DNA fragments of what
sizes?
Which lane(s) of the gel shows this expected DNA fragmentation pattern?
c. If you want to clone a piece of DNA into the MCS (labeled in blue), which restriction enzyme(s) could
you use?
d. After digestion of the plasmid and the DNA insert with this restriction enzyme, what enzymatic
reaction is necessary to generate the desired recombinant plasmid DNA?
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