Uncut - DNA Fragments produced B2 Sspl Enzyme(s) EcoR1 Smal used - 5.0 plasmid, how many times does it cut? Explain briefly (1 pt) d. Do you think Smal cuts the plasmid, how many times does it cut? Explain briefly (2 pt) To determine the restriction map of plasmid RK21 DNA, you perform the following complete digests with Xhol, Smal and EcoRI. You run the digested DNA on an agarose gel, and obtain the following fragments (all sizes are in kilobases) 2.9 2.1 B1 Xhol -C Smal Xhol EcoRI + Smal 5.0 a. Explain briefly the two observed bands of the uncut DNA sample. (2 pt) 2.3 2.1 0.6 b. Do you think Sspl cuts the plasmid, how many times does it cut? Explain briefly (1 pt) c. Do you think Xhol cuts the EcoRI + Xhol Smal + Xhol 4.0 1.0 Using the above information & the incomplete map below, answer the following regarding the restriction map of plasmid RK21 DNA. a. What is the size of plasmid RK21 in base pairs? (1 pt) b. Which of the above restriction enzymes is A? (1 pt) c. Which of the above restriction enzymes is B? (1 pt) d. Which of the above restriction enzymes is C? (1 pt) e. What is the distance in base pairs between A & B1? (1 pt) f. What is the distance in base pairs between C & B1? (1 pt) g. What is the distance in base pairs between C & B2? (1 pt) 2.9 1.7 0.4 h. What is the distance in base pairs between A & B2? (1 pt) EcoRI + Smal + Xhol 2.3 1.7 0.6 0.4

Biochemistry

6th Edition

ISBN:9781305577206

Author:Reginald H. Garrett, Charles M. Grisham

Publisher:Reginald H. Garrett, Charles M. Grisham

Chapter28: Dna Metabolism: Replication, Recombination, And Repair

Section: Chapter Questions

Problem 1P: Semiconservative or Conservative DNA Replication If 15N-Iabeled E. coli DNA has a density of 1.724...

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You have discovered a new plasmid RK21 in a unique bacterial community. As a first step toward
understanding this plasmid, you digest the plasmid with three restriction enzymes: SspI, XhoI and
SmaI. You run the digested plasmid DNA on an agarose gel, along with an uncut sample of the
RK21 plasmid DNA as a control.
Unfortunately you forget to load a DNA ladder, and obtain the following results. Assume
complete digestion of all samples or all the digests worked completely

Uncut
-
DNA
Fragments
produced
B2
Sspl
Enzyme(s) EcoR1 Smal
used
-
5.0
plasmid, how many times does it cut? Explain briefly (1 pt)
d. Do you think Smal cuts the plasmid, how many times does it cut? Explain briefly (2 pt)
To determine the restriction map of plasmid RK21 DNA, you perform the following complete digests
with Xhol, Smal and EcoRI. You run the digested DNA on an agarose gel, and obtain the following
fragments (all sizes are in kilobases)
2.9
2.1
B1
Xhol
-C
Smal
Xhol EcoRI + Smal
5.0
a. Explain briefly the two
observed bands of the uncut
DNA sample. (2 pt)
2.3
2.1
0.6
b. Do you think Sspl cuts the
plasmid, how many times does
it cut? Explain briefly (1 pt)
c. Do you think Xhol cuts the
EcoRI + Xhol Smal + Xhol
4.0
1.0
Using the above information & the incomplete map below, answer the following regarding the
restriction map of plasmid RK21 DNA.
a. What is the size of plasmid RK21 in base pairs? (1 pt)
b. Which of the above restriction enzymes is A? (1 pt)
c. Which of the above restriction enzymes is B? (1 pt)
d. Which of the above restriction enzymes is C? (1 pt)
e.
What is the distance in base pairs between A & B1? (1
pt)
f. What is the distance in base pairs between C & B1? (1
pt)
g.
What is the distance in base pairs between C & B2? (1 pt)
2.9
1.7
0.4
h.
What is the distance in base pairs between A & B2? (1 pt)
EcoRI +
Smal+
Xhol
2.3
1.7
0.6
0.4

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