Laboratory Operations Toolbox Teacher Guide
Biotechnology in Action
Description
This Biotechnology unit is designed to complement the Toolbox 5 series of
laboratory-based units and the scenario chosen follows and adds to each of the units
in Toolbox 5. The following table demonstrates the scenario’s relationship to the
competency units.
Unit relationship to competency
Competency
Scenario section in Toolbox
Step 1. Introduction to the Scenario
1. COM500A Provide information to
customers
Step 2. Communication with Customer
2. ORG500A Schedule laboratory work
for a small team
Step 3. Planning for extraction of DNA
3. MAIN500A Maintain and control
stocks
Step 4. Maintaining the Biotechnology
Laboratory
4. TEST506A Apply spectrometric
techniques
Step 5. Extraction and Quantitation of DNA
from Lizard Skin
5. TEST500A Calibrate and maintain
instruments
Step 6. Calibration and Setting up of Equipment
6. PMLTEST507A Apply
Chromatographic and Electrophoretic
Techniques
Step 7. Running the Gel
1. COM500A Provide information to
customers
Step 8. Communication of Results.
Please also note that this biotechnology unit has the following features and does not
purport to be a comprehensive introduction to biotechnology.
Features of this unit:

A brief introduction to biotechnology in general

A brief introduction to some of the concepts of biotechnology

An exercise to start the student thinking about biotechnology, and

A ‘taster’ only, as there is no in-depth material in this unit.
Depending on circumstances you may wish to use this unit as an introduction to
biotechnology and/or add additional materials specific to requirements to ‘flesh out’
the unit.
Teacher Guide Biotechnology in Action
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Laboratory Operations Toolbox Teacher Guide
Step: 3
Forum: Extraction/purification and amplification of DNA
In this step, learners will participate in a discussion forum about the
extraction/purification and amplification of DNA. Before participating in the
discussion forum the learner needs to access some information.
The learner should go to the ROCHE website and obtain information on the
extraction of DNA from tissues (eg. go to the World Wide Web Catalogue and choose
Web Visit – ROCHE from the list provided).
Other sites can then be investigated using a search engine term such as ‘DNA
extraction’ to obtain further information, especially alternative methods of DNA
extraction. The best sites to look at are company-based (suppliers to the
biotechnology industry such as ROCHE) or research based (e.g. university or
biotechnology company research information). Look for at least one method of PCR
amplification.
Forum Discussion
When you have completed your Internet search go to the Forum and choose the
topic Extraction/purification and amplification of DNA.
Tutor Note
The discussion could be initiated with questions such as: Why is DNA extracted from
cells and tissues? How is it extracted? How is purity determined? How is the DNA
amplified?
In addressing these questions, the learners should discuss use of different reagents
during the extraction and purification process. When discussing purity, an
explanation of how protein and RNA contamination is detected should be included.
On the question of amplification, an explanation of PCR and the importance of
temperature program can be raised. It should also include some discussion of the
absolute requirement for cleanliness during PCR.
Teacher Guide Biotechnology in Action
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Laboratory Operations Toolbox Teacher Guide
Step: 3
Activity: Assignment - DNA Extraction, Purification and Amplification
Questions and Answers
Answer the following questions based on your Internet website search for DNA
Extraction/Purification and PCR.
1. What is DNA extraction/purification and why is it used?
DNA extraction/purification is the method employed to extract DNA from cellular
or tissue samples and is used to prepare adequate amounts of DNA of high
enough purity for subsequent analysis of the DNA.
2. List one method for DNA extraction sourced from a website, such as the ROCHE
site. You should present this as a flowchart listing each of the steps. There is no
need to explain the steps.
There are a number of methods listed on this site. This assignment is designed
to make the student aware of DNA extraction/purification as the first steps in DNA
analysis BUT not to make the student an expert in this technique. Look for an
answer the clearly lists all the steps in the chosen technique.
3. List one other method of DNA extraction and list the website from which it was
obtained. Again, use a flowchart to outline the steps.
There are innumerable methods listed on the Internet. This assignment is
designed to make the student aware of DNA extraction/purification as the first
step in DNA analysis BUT not to make the student an expert in this technique.
Look for an answer the clearly lists all the steps in the chosen technique.
4. Which of these methods was the better one and why.
Look for a clear indication of the better method. Reasons include:
 cheaper
 safer reagents
 less steps - less complicated
 less time
 better yield (quantity)
 better quality.
5. List one method of PCR and list the website from which it was obtained. Again,
use a flowchart to outline the steps.
There are innumerable methods listed on the Internet. This assignment is
designed to make the student aware of PCR as the amplification step in DNA
analysis BUT not to make the student an expert in this technique. Look for an
answer the clearly lists all the steps in the chosen technique.
Teacher Guide Biotechnology in Action
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Laboratory Operations Toolbox Teacher Guide
Step: 6
Activity: Assignment - Agarose Gel Electrophoresis
Questions and Answers
Answer the following questions using SOP: Agarose Gel Electrophoresis as a
guide.
1. When setting up the agarose gel apparatus, including pouring the gel, name
three important factors that have a direct bearing of the success of the run. In
this case success means a gel with clearly separated and non-distorted bands.
 Correct set up of apparatus

No leaks

Correct strength of agarose


Samples placed into well correctly
Correct buffers used

Correct running conditions - voltage and time

Agarose prepared correctly - fully dissolved


Agarose allowed to set fully
Tape removed from ends of casting plate

Gel stain added.
2. Can you calibrate the gel apparatus itself? Explain your answer.
No, because there are no mechanisms for calibration and nothing that can be
adjusted.
3. Can you calibrate the gel itself to account for small variations between separate
gels? Explain your answer.
Yes, the use of a DNA marker allows calibration of the run, as the sizes of the
DNA fragments in the samples are determined from the known sizes of the DNA
fragments in the ladder.
4. Name three safety concerns when doing agarose gel electrophoresis.

Ethidium bromide is a mutagen

Biosafety issues arising from the DNA samples

Electricity


Burns form molten agar
General chemical safety from buffer constituents

UV light damage to eyes and skin.
5. Given what you know about the basics of electrophoresis, why do you think that
the concentration of agarose may be varied from 1 - 3%?
The concentration of agarose is adjusted to suit the size of the fragments being
analysed. Smaller DNA fragments require a higher strength agarose for effective
resolution and larger DNA fragments require a lower strength agarose.
Teacher Guide Biotechnology in Action
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Laboratory Operations Toolbox Teacher Guide
Step: 8
Question and Answers: Correctly filled in form
Laboratory Test Sheet - Lizard DNA Samples
Date: x/1/2004
Customer Sample Numbers: Lizard 120, Lizard 121, Lizard
122, Lizard 123, George, Fred
Sample Description: Lizard skin
Sample Type: DNA
Sample Volume: N/A - sheet of skin
Customer Sample Number
Integrity of Sample
Sample Accepted/Rejected
for testing.
Lizard 120
Lizard 121
Lizard 122
Lizard 123
George
Fred
All samples are intact and
suitable for analysis.
All samples accepted, none
rejected.
Client Details:
Australian Reptiles Pty Ltd
5 Big River Road
SimuTown CIV 0313
Contact:
Dr Jake Crawford, Head breeder
9551-2106
Integrity of Samples: Details of any rejected samples are listed here:
..................................................
..................................................
..................................................
Tests Required/Results (Bold and underline testing required):

Parentage

Sex Determination


Sequencing
Other
TEST Results:
Sample Identification
Lizard 120
Lizard 121
Lizard 122
Lizard 123
Test Result - Paternity
George
George
George
George
Test Result - Sex
Male
Female
Female
Female
Checked by: Student name
Teacher Guide Biotechnology in Action
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