Bi 12.

02 Biotechnology for Everyone, Laboratory

Exercise 6: DNA Extraction and Agarose Gel Electrophoresis

WORKSHEET

Names of Members: _______________ , _______________

Results/Discussion:

I. Virtual laboratory #2: Take a screenshot of your final agarose gel, with the estimated
molecular weights for each resulting DNA band.

II. Analyze the agarose gels below and answer the questions that follow.

1. Agarose gel #1:

Note: + and – represent the positive and negative controls, while the DNA samples
tested are in lanes A, B, and C.

a. Which of the following samples (A/B/C) has the smallest molecular weight? _______
b. What is the estimated molecular weight of Sample C? _______
c. Identify two errors in the gel.

Department of Biology, School of Science and Engineering

Ateneo de Manila University
Bi 12.02 Biotechnology for Everyone, Laboratory
2. Agarose gel #2:

a. Identify the error in the gel, and why is this error crucial in gel analysis?

3. Agarose gel #3:

a. Student Y performed agarose gel electrophoresis to check the molecular weight of his
genomic DNA isolated from bioluminescent bacteria. Upon exposure to UV however,
he was unable to see a single band on the gel. What might have caused this to occur?

Department of Biology, School of Science and Engineering

Ateneo de Manila University
Bi 12.02 Biotechnology for Everyone, Laboratory
III. Answer the following questions. Limit your answer for each question to a maximum of five
(5) sentences only.

1. What is the purpose of DNA extraction? How can this be applied different fields of
Biotechnology?

2. What is contained in a lysis buffer? Why is it the first step in DNA extraction?

3. What is the purpose of adding isopropyl alcohol to the solution in the process of DNA
extraction?

4. What are the components of the loading dye/buffer? Identify the uses of each component.

5. Why is it important that the agarose is exposed to ethidium bromide, which is either added to
the gel itself or mixed to the staining solution? Explain the principle behind its use.

References:

Department of Biology, School of Science and Engineering

Ateneo de Manila University