10 Advanced Science and Engineering Name: ________________________

Practical Assessment: Biotechnology

This assessment will take place over three lessons and be broken down as follows:
 Lesson One - Part A: Gel Electrophoresis – There has been a number of serious
crimes undertaken in the community over the past few weeks and fortunately
forensics have found a small portion of DNA at the latest crime scene. As a result,
local detectives have managed to secure DNA from two possible suspects. Your task
will be to use Gel Electrophoresis to determine who the perpetrator is.
 Lesson Two – Part B: Staining of Gels – You will complete the staining process in
order to determine the results of your electrophoresis lab. You will be assessed on
the quality of your results as well as your detailed observations. You will also have to
make your decision about who you believe the perpetrator of these crimes is.
 Lesson Three – Validation Test - You will complete a 30-minute validation test
covering Gel Electrophoresis and associated terminology.

Lesson One - Part A: Gel Electrophoresis

Experiment Overview

Experimental Procedure:

DONE PRIOR TO CLASS: (by Lab Technicians)

 1. DILUTE concentrated (50X) buffer with distilled water to create 1X buffer.
2. MIX agarose powder with 1X buffer in a 250 ml flask.
3. DISSOLVE agarose powder by boiling the solution. MICROWAVE the solution on
high for 1 minute. Carefully REMOVE the flask from the microwave and MIX by
swirling the flask. Continue to HEAT the solution in 15-second bursts until the
agarose is completely dissolved (the solution should be clear like water).
4. COOL agarose to 60° C with careful swirling to promote even dissipation of heat.
5. While agarose is cooling, SEAL the ends of the gel-casting tray with the rubber end
caps. PLACE the well template (comb) in the appropriate notch.
6. POUR the cooled agarose solution into the prepared gel-casting tray. The gel
should thoroughly solidify within 20 minutes. The gel will stiffen and become less
transparent as it solidifies.
7. REMOVE end caps and comb. Take particular care when removing the comb to
prevent damage to the wells.
8. PLACE gel (on the tray) into electrophoresis chamber. COVER the gel with 1X
electrophoresis buffer. The gel should be completely submerged.

DONE IN CLASS:

9. LOAD the entire sample (35-38 μL) into the well in the order indicated by Table 1
(below)

10. PLACE safety cover. CHECK that the gel is properly oriented. Remember, the DNA
samples will migrate toward the positive (red) electrode.
11. CONNECT leads to the power source and PERFORM electrophoresis.

TO BE DONE AFTER CLASS (by Lab Technicians)

12. After electrophoresis is complete, REMOVE the gel and casting tray from the
electrophoresis chamber and proceed to STAINING the agarose gel.

Lesson Two - Part B: Staining of Gels

Experiment Overview

Experimental Procedure:

Staining was completed by the lab technicians overnight and your results are now ready to
be analysed. In order to analyse your gel you will be required to:

1.
Observations:
1. Using your transparency as a guide, use the box below to draw the results of
running the Gel Electrophoresis.
(2 marks)

2. Identify which of the suspects is the perpetrator. Explain your answer.

(2 marks)

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3. Explain if these DNA samples could have been distinguished from one another if
only Enzyme 1 had been used?
(2 marks)

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4. Explain if the DNA of victim of the crime should have been run at the same time?
(2 marks)

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