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An NCBE / Unilever educational guide

Introduction | Equipment | Acknowledgements | Copyright

to the reader
Work with DNA is central to many, if not most, developments
in modern biotechnology. There is growing public awareness
of DNA technologies, their possible applications and wider
implications. However, much of the essential debate about
current genetics has generated more heat than light.
Fortunately, the basic science upon which DNA technologies
are founded features in nearly every school examination
syllabus in biology or science. This booklet is intended
primarily for post-16 students of biology and their teachers.
The practical exercises described here provide an introduction
to some of the classical techniques of molecular biology in a
form suitable for the school laboratory. For reasons of safety
and expense, some the work in this booklet is not particularly
suited to open-ended practical investigations, but some ideas
that may provide starting points for additional work of your
own are given on pages 3637. More ideas are provided in
other NCBE publications such as: The Lambda Protocol;

Investigating Plant DNA and The Transformer Protocol. The


latter publication also provides an introduction to ethical,
social and other issues raised by DNA technology. All of
these publications, the other practical guides in this series,
and supplementary resources are available from the NCBEs
Web site:
http://www.ncbe.reading.ac.uk
An adequate treatment of the wider issues raised by DNA
technology (particularly those associated with human
genetics, environmental concerns or food biotechnology)
cannot be given in this slim booklet, nor can it take the place
of the many excellent school textbooks covering basic
biochemistry and molecular biology. It aims merely to provide
sufficient information for you to understand the practical
exercises. References to other materials are provided
throughout and on the back cover. The NCBEs Web site
offers a more comprehensive list.

equipment and materials


In this booklet we have tried to describe practical protocols
that are relatively inexpensive and require little specialist
equipment.
The NCBE endeavours to develop and supply at low cost
several of the items required, such as electrophoresis
apparatus, restriction enzymes, plasmids and so on.
Because the details of suppliers and their products can change,
we have decided not to list them in this booklet. Instead, a
supplement giving up-to-date information is available from
the NCBEs Web site, or it can be obtained on request from
the NCBE. Our address is given on the back page.

Some of the investigations


described here require a
high-speed microcentrifuge.
An inexpensive, awardwinning microcentrifuge
for school use has been
developed by our friend
Professor John Cave of
Middlesex University and
the Technology Enhancement
Programme (TEP), with help
from Science and Plants for
Schools (SAPS) and the NCBE.

acknowledgements
The production of this booklet was very kindly supported
by Unilever plc. The contents are entirely the responsibility
of the NCBE however. Unilevers only conditions in
sponsoring this booklet were that we made the publication
freely available via the Internet, and that we consulted our
colleague, Dr Malcolm Thomas, in the Department of
Education at the University of Aberystwyth. Malcolm made
many helpful comments, for which we are grateful. Marjorie
Smith of Dollar Academy was very supportive too, giving us
tips for making this booklet more palatable to those in
Scotland. The molecular structure data for several of the
figures came from the Protein Data Bank and the Nucleic
Acid Database; accession numbers are given by the figures.
Any errors in this booklet are the authors, of course.
2| ILLUMINATING DNA | Version 1.0 | June 2000

COPYRIGHT
The material in this booklet is copyright.
The author and illustrator, Dean Madden, has asserted his moral
right to be identified as the copyright holder under Section 77 of
the Copyright, Designs and Patents Act, UK (1988).

PHOTOCOPYING
Photocopies of this booklet, or selections from it may be made
for educational use, provided that the copies are distributed freeof-charge or at the cost of reproduction, and the author of the
materials is credited and identified as the copyright holder.

contents
04 DNA: molecule of the century
DNA, genes and chromosomes | The genetic code and protein synthesis | Enzymes, precise
molecular tools | Bacterial transformation | Genetically modified plants and animals | The
polymerase chain reaction | Marker genes and gene regulation | DNA gel electrophoresis
| Gene mapping | DNA sequencing | Using genetic data

14 Safety guidelines
Good microbiological laboratory practice

18 Practical activities
Modelling DNA
Physical and computer models are useful for studying molecular structures

18

Extracting DNA
Simple extraction of DNA from onions or fish sperm

20

DNA quantification
Diphenylamine gives a blue-coloured complex with deoxyribose
that can be used to measure the amount of DNA in a sample

22

E. coli makes the enzyme -galactosidase but only when induced to

24

-galactosidase induction

Extraction of plasmid DNA


Plasmid DNA can be extracted from bacteria and run on an electrophoresis gel

26

Bacterial transformation
Prove that DNA is the genetic material by transforming bacteria

28

Restriction and ligation


Restriction enzymes and DNA ligase can be used to cut and paste DNA

30

Restriction site mapping


Find out where the restriction sites are in phage Lambdas DNA

32

Amplifying Lambda DNA


The polymerase chain reaction has been likened to a genetic photocopier;
use it here to amplify a fragment of Lambdas DNA

34

36 Ideas for investigations


38 Running gels | Units of measurement
39 Staining DNA on electrophoresis gels
40 Recipes
42 Glossary
44 Other sources of information
www.ncbe.reading.ac.uk |3