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CIE A Level Biology

Revision Notes

A Level Biology CIE Revision Notes

19. Genetic Technology
19.1 Principles of Genetic Technology Microarrays

Syllabus Edition
First teaching 2023
First exams 2025

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Microarrays
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Microarrays
Microarrays are laboratory tools used to
detect the expression of thousands of
genes at the same time and to identify
the genes present in an organism’s
genome
Microarrays are used in di!erent ways,
e.g.
Pathology: comparison between
healthy cells and diseased cells to
find the characteristics of the
disease
Biotechnology (e.g. in agriculture to
identify insect pests)
Crime (e.g. forensic analysis)
As large numbers of genes can be
studied in a short period of time,
microarrays have been very valuable to
scientists
The microarray consists of a small (usually
2cm2) piece of glass, plastic or silicon
(also known as chips) that have probes
attached to a spot (called a gene spot) in
a grid pattern
There can be 10 000 or more spots
per cm2
Probes are short lengths of single-
stranded DNA (oligonucleotides) or RNA
which are synthesised to be
complementary for a specific base
sequence (this sequence depends on
the purpose of the microarray)

When a microarray is used to analyse

genomes:

DNA is collected from the species going

to be compared
Restriction enzymes are used to cut the
DNA into fragments
These fragments are denatured to create
single-stranded DNA molecules
These DNA fragments are labelled using
fluorescent tags (the fragments from the
di!erent sources are tagged in di!erent
colours, usually red and green)
Once these fragments are mixed
together they are allowed to hybridise
with the probes on the microarray
After a set period of time, any DNA that
did not hybridise with the probes is
washed o!
The microarray is then examined using
ultraviolet light (which causes the tags to
fluoresce) or scanned (colours are
detected by the computer and the
information is analysed and stored)
The presence of the colour indicates
where hybridisation has occurred, as the
DNA fragment is complementary to the
probe
If red and green fluorescent spots
appear then only one species of DNA
has hybridised, however, if the spot
is yellow then both species have
hybridised with that DNA fragment,
which suggests that both species
have that gene in common
If a spot lacks colour that indicates the
gene is not present in either species

When genes are being expressed or are

in their active state, many copies of
mRNA are produced by transcription
The corresponding proteins are then
produced from these mRNAs during
translation
Thus scientists can indirectly, by
assessing the quantity of mRNAs,
determine which genes are being
expressed in the cells
Microarrays can be used to detect
whether a gene is being expressed (a
method used to research cancerous vs
non-cancerous cells) by detecting the
quantity of mRNA present
To compare which genes are being
expressed using microarrays the
following steps occur:
mRNA is collected from both types
of cells and reverse transcriptase is
used to convert mRNA into cDNA
PCR may be used to increase the
quantity of cDNA (this occurs for all
samples to remain proportional so a
comparison can be made when
analysis occurs)
Fluorescent tags are added to the
cDNA
The cDNA is then denatured to
produce single-stranded DNA
The single-stranded DNA molecules
are allowed to hybridise with the
probes on the microarray
When the ultraviolet light is shone on
the microarray the spots that
fluoresce indicate that the gene was
transcribed (expressed) and the
intensity of the light emitting from
the spots indicates the quantity of
mRNA produced (i.e. how active the
gene is)
If the light being emitted is of high
intensity then many mRNA were
present, while a low intensity
emission indicates few mRNA are
present

The Process of Microarrays Diagram

The process of using microarrays to

compare the expression of genes in
cancerous and non-cancerous cells. The
colours on the microarray identify which
genes are active in cancerous cells
compared to normal cells

Exam Tip
The colours of the fluorescent tags (red,
green and yellow) indicate whether a gene is
present whereas the intensity of light
emitted indicates the level of gene
expression (the more light, the more the
gene was being expressed) and this relates
to the quantity of mRNA present.