Name: Date:
Problem set II. Genetic Engineering Worksheet
1. Below are simplified representations of DNA sequences from three different sources—labeled Person
A, Person B, and Person C. Specific marker points (shown as arrows) indicate spots where molecular
scissors (enzymes) would cut the strands.
Your task: Analyze each sequence to determine how many pieces (fragments) each DNA strand would
be divided into. Estimate the relative size of each resulting segment. Sketch what the final result
would look like if you ran this through a gel-based separation method (think about how pieces of
different sizes would appear). Note: Focus on interpreting the cut points and how they affect the
number and size of the pieces. Use logic and pattern recognition to complete the diagram.
2. Show bands to indicate the location of the DNA fragments that would be formed in a gel
electrophoresis (from each segment above) on the gel below. Label the side of the gel with numbers
to indicate size of the DNA fragments in each band. (Bands with the same number of nucleotides should
be shown at the same distance from the wells.) Also, indicate which side of the gel box would be
positively charged and which would be negatively charged.
+ or –
A B C
Circle One
Gel
+ or –
Circle One
1
Use the following figure to answer questions 8-11. The plasmid pGEN101 shown was treated with
various mixtures of restriction enzymes. The electrophoresis gel shows the results of each of those
digestions.
3. Which lane (I to V) represents the fragments produced using Bam HI only? ___________
4. Which lane represents the fragments produced using Eco RI only? ____________________
5. Which lane represents the fragments produced when the plasmid was cut with both Eco RI and
Bam HI? ________________
6. The segment of DNA pictured below has been cut with specific enzymes which create fragments A,
B, and C.
Which of the following gels produced by electrophoresis would represent the separation and
identity of these fragments?
A. C.
B. D.
2
The following electrophoresis gel shows fragments obtained from a plasmid digested with the
restriction enzymes X and Y. The fragments cut by enzyme X alone are in Lane 1; the fragments cut by
enzyme Y alone are in Lane 2; and the double digest using both enzymes X and Y is in Lane 3.
7. You're provided with a circular DNA diagram (plasmid) that has three marked cut points,
shown as slashes through the circle. Each slash represents where a specific enzyme—either
Enzyme X or Enzyme Y—makes a cut. Next to each region of the plasmid, you'll see a number that
represents the length of DNA (in nucleotides) between cuts. A gel image showing the result of
these cuts has also been provided. Carefully study both the plasmid map and the gel image. Decide
which cut(s) were made by Enzyme X and which were made by Enzyme Y. Use the fragment sizes
and number of bands in the gel to help guide your reasoning. Think about how circular DNA
behaves when it's cut once, twice, or three times, and how the resulting fragment sizes would look
in a gel.
Determine the cuts 1, 2, and 3 would be made by which enzymes, respectively?
a. X, Y, Y b. Y, X, Y c. Y, Y, X d. X, X, Y