11 Jun Lab 10 Biomolecular – Restriction Endonucleases Materials
Question
Lab 10. Biomolecular
Biomolecular
Techniques Techniques eScience ref #: Lab 14 Experiment 2: Restriction Endonucleases Materials
12 Pop-it® Beads (First Color) Note: The Pop-it® beads bag contains randomly 9 Pop-it® Beads (Second Color) colored beads. Each bag contains at least one color with 12 beads, one color with nine beads, one 8 Pop-it® Beads (Third Color) color with eight beads, and one color with six ® 6 Pop-it Beads (Fourth Color) beads. However, you will have to count the beads *Colored Pencils to determine which color you should use for each
of the four categories (listed in the left-hand column of Table 1). *You Must Provide Figure 8: Sample nucleotide sequence. Your color sequence may vary. Note the directionality on the DNA strand. Table 1: Enzyme Analysis
Cuts Between… Fragment Sizes Gel Banding Pattern Enzyme 1
Enzyme 2
Enzyme 3
Enzyme 4
Enzymes 2 and 3 (see above
combinations) Procedure
1. Use the Figure 8 as a reference to construct a strand of Pop-it® beads with the same color pattern, although different colors may be used (Figure 8).
2. Use ‘enzyme 1’ to cut the strand of DNA (Pop-it® beads) based on the ligation information provided in Table 1. Biomolecular Techniques
Note: Remember, your DNA strand color scheme may vary, but the pattern should be the same
as is displayed in Figure 8. For example, Enzyme 1 cuts between 2 blue beads in Table 1, but it
may correspond to green, red, or yellow beads in your experiment (depending on which colors
you selected). It does not matter which color you assign to each category, but be sure to keep the
same pattern used created in Step 1.
3. Use colored pencils to draw the fragment sizes in the appropriate cell of Table 1.
4. Line up the fragments as they would separate if run through an electrophoresis gel. Use the
colored pencils to draw them in the appropriate cell of Table 1.
5. Repeat this procedure for each enzyme.
Note: The bottom row titled “Enzyme 2 and 3” means that first, Enzyme 2 fragments the
DNA, then Enzyme 3 cuts the fragments made by Enzyme 2).
Post-Lab Assessment 1. How can DNA be fragmented into very specific sections?
2. Where do restriction enzymes come from? What is their function in nature?
3. How do molecules of varying sizes separate in electrophoresis? What is the purpose of the
gel? What about the electricity?
4. Investigate one way in which electrophoresis is used in medicine today. Write one
or two paragraphs to explain your findings.
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