Академический Документы
Профессиональный Документы
Культура Документы
DNA Fingerprinting
Lesson Objectives
Describe the uses for DNA Fingerprinting
Explain the process of DNA Fingerprinting
Discuss the ethics behind DNA
Figerprinting
Restriction Enzymes
The DNA sample is treated with
Restriction Enzymes
These Enzymes cut DNA at specific sites
dictated by the sequence at that site
The DNA is cut up into a series of lengths
These lengths vary in size between
individuals (even animals!)
Gel Electrophoresis
Gel Electrophoresis is a technique used to
separate molecules by their size (and
even by charge)
It can be used to separate different sizes
of lengths of DNA
How it works
A gel is made from a chemical called agarose
(made from seaweed!)
Samples of unknown DNA and DNA of known
lengths are loaded at one end of the gel
An electrical current is passed along the gel
As DNA is mainly negative it is attracted to the
positive electrode
The smaller the DNA fragment the quicker and
further it travels along the gel
Adding samples
Electrophoresis
Southern Blotting
The fragments of DNA within the gel are
transferred to a nylon membrane and washed
over with a DNA probe that binds to the repeated
sequence
The membrane is then placed in a bag and
placed on a photographic film which is exposed
where the radioactive probes are attached
The resulting pattern of bands is called the DNA
fingerprint
A single band occurs where the maternal and
paternal chromosomes have the same number
of satellite repeats, if not there will be two bands
Summary
Automation
This shows the result of a modern automated
technique using fluorescent DNA to make a
computer generated graph
Amelogenin
The genes for amelogenin can be used in sex determination of
samples from unknown human origin through the Polymerase Chain
Reaction (PCR).
Using primers specific for intron 1 of the gene, the gene sequence
for the intron can be amplified. The X chromosome gene, AMELX,
gives rise to a 106 bp amplification product (amplicon) and the
Y chromosome gene, AMELY, a 112 bp amplicon. Hence, the
AMELX contains a 6 bp deletion in the intron 1.
When the amplicons are run on an agarose gel, samples from male
sources (XY) will show two bands on an agarose gel (one for the
106 bp fragment and one for the 112 bp fragment), while females
(XX) will show only one band.
Thus, this process allows for sex determination of unknown
samples.
Interpretation
In the UK ten loci are copied during DNA
fingerprinting
In the USA thirteen loci (they call them
alleles) are copied, why?
Because the USA has a larger population
and so there is more chance of a
similar/identical DNA fingerprint
Even so this is still very unlikely
Matching Loci
To match a sample of DNA all ten loci
must be of the same length!
If even one is different there is no match
The defendant stated that the blood on his clothing was his.
A mother
B male 1
C male 2
D child
E standards
DNA Databases
In England and Wales, anyone arrested on suspicion of a
recordable offence must submit a DNA sample to the database,
which is then kept on permanent record.
In Scotland, the law is different and most people are removed from
the database if they are acquitted. In Sweden, only criminals who
have spent more than two years in prison are recorded.
In Norway and Germany, court orders are required, and are only
available, respectively, for serious offenders and for those convicted
of certain offences and likely to reoffend.
All 50 states in the USA keep profiles of violent offenders, and a few
keep profiles of suspects.
Portugal has plans to introduce a DNA database of its entire
population