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Principles of Genetic Technology _ Cambridge (CIE) A Level Biology Revision Notes 2023

Microarrays are laboratory tools used to analyze gene expression and identify genes in an organism's genome, with applications in pathology, biotechnology, and forensic analysis. The process involves hybridizing labeled DNA fragments to probes on a microarray, with fluorescent tags indicating gene presence and expression levels. By assessing the quantity of mRNA, scientists can determine which genes are active, particularly in comparing cancerous and non-cancerous cells.

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9 views4 pages

Principles of Genetic Technology _ Cambridge (CIE) A Level Biology Revision Notes 2023

Microarrays are laboratory tools used to analyze gene expression and identify genes in an organism's genome, with applications in pathology, biotechnology, and forensic analysis. The process involves hybridizing labeled DNA fragments to probes on a microarray, with fluorescent tags indicating gene presence and expression levels. By assessing the quantity of mRNA, scientists can determine which genes are active, particularly in comparing cancerous and non-cancerous cells.

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Microarrays
Your notes
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 different 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 different sources
are tagged in different 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 off
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
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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 Your notes
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

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Your notes

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Your notes

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

Examiner Tips and Tricks


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.

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