PCR Information and Resources for Teachers
PCR is a tool which expands the applications
of other molecular biology tools by reducing the required initial
nucleic acid sample size and the time required to generate much
larger quantities of DNA for analysis.
DNA or RNA samples which are too small to be analyzed by more
traditional molecular techniques are easily amplified with PCR so
that information about the source and content of the nucleic acid
are generated in a short time.
Sample size is often a limiting factor in nucleic acid
analysis. For example,
samples from crime scenes tend to be collected from a few drops of
blood, a few hairs or a small bit of skin . Once these small samples are amplified by PCR, other
molecular biology tools which require greater quantities of DNA can
be used to analyze the DNA. This
broadens the areas of application for molecular biology.
Clinical Applications
PCR is useful in establishing the unambiguous
identity of a sample. A
“sample” can be any source of DNA or RNA from an organism to a
few cells. Samples are
usually small amounts of blood or other body fluids, or a small
collection of cells such as a cheek swab, a bit of bone or some
hair.
Medical
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Genetic diseases can be identified pre- or post-natally.
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Cancers can be identified and classified.
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Identifying bacterial or viral strains for proper treatment
or epidemiological studies.
Food Science
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Samples can be tested for genetic engineering traits.
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Bacterial contaminants and the source of contamination can be
quickly identified.
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Pedigrees and traits of valuable animal food stocks can be
verified.
Forensics
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Identification of corpses or body parts.
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Analysis of pathogens
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PCR is a valuable tool in criminal investigation where it is
used to eliminate possible suspects as well as link suspects to the
scene of the crime either through a sample (blood, human hair, skin,
semen) left at the scene or circumstantial links (pet hair, plant
parts).
Relationships
PCR can also be used to establish relationships
between samples, as in the following three examples:
Paternity
This method also demonstrates maternity, but
maternity is less often in question.
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Identifying familial relationships in organisms such as
wolves or chimpanzees.
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Verifying pedigrees of valuable stock such as horses, cows
and dogs.
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Providing evidence for paternity in custody disputes.
Evolution
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archaeological analysis of samples from sites
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Developing relationships based on molecular phylogeny.
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Studying extinct species based on museum specimens.
Ecology
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Validating conservation efforts by tracking rare wildlife or
individual organisms.
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Clearly defining unique subgroups within populations or
species
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Defining biodiversity by identifying unique species.
Useful web sites
Forensics
(http://www.ornl.gov/TechResources/Human_Genome/elsi/forensics.html)
This is a page from the Human Genome Project
and has information on forensics.
This page links to a page with several very interesting cases
of forensic identification many of which have current personal and
political ramifications.
DNA testing
Private individuals may contract with a
number of companies offering DNA testing.
These companies tout their services not only for criminal
investigations, but also for paternity, pre-natal testing, and
identity establishment for estate claims and immigration purposes.
There is also a market for veterinary uses of DNA testing
including pedigree, identity and sexing birds.
Search for “DNA Typing” on the internet to investigate
DNA testing companies.
DNA testing raises a variety of ethical
issues, specifically privacy issues which students will be faced
with in their lifetimes. This
site has an excellent essay on DNA testing issues and asks a number
of ethical questions as well:
http://www.accessexcellence.org/AE/AEPC/WWC/1992/gen_screen1.html)
1) Since non-invasive samples may be used for DNA testing (such
as a toothbrush or hair), people can be tested without their
knowledge or consent. Should
this be legal?
2)
The military now takes a blood sample from each person
entering the military to aid in identifying bodies or body parts.
The claim is that there will never be another “unknown
soldier”. Many states
are compiling criminal genetic records as well.
This helps police identify repeat offenders or link a suspect
in one case to another case. These
records could potentially be used in a number of ways such as
academic and medical studies where general information would be
collected with the genetic information, but not personal
identifiers, such as names or social security numbers.
Other groups with profit in mind such as insurance companies
or drug manufacturers might wish to access genetic information as
well. Should genetic records be maintained, and if so, by whom?
Who should have access to this information?
3)
What limitations should be placed on pre-natal testing?
Is it legitimate to test for gender, potential for developing
adult onset diseases (Huntington’s, aggressive breast cancer) or
fatal genetic flaws such as Lesch-Nyhan syndrome, which results in a
short, painful life characterized by severe retardation, violence,
and self-mutilation? What
about non-fatal diseases or syndromes such as Down’s syndrome or
galactosemia which require life long care and can generate other
medical issues? What
about diseases that can be treated, but at great cost, and with a
probable decrease in life span such as Gaucher’s?
4)
DNA testing can potentially lead to the development of a
“genetic underclass”, those who cannot access DNA testing and so
continue to produce children with genetic defects. What ethical considerations arise in this situation?
What are the ramifications for these people economically such
as lost work time, higher medical bills and special care costs?
What role will insurance companies play in these issues?
Will DNA testing be mandatory for health insurance, and if
so, to what information will insurance companies be allowed access?
5)
Many medical advances have changed previously deadly diseases
into diseases that can be survived or managed for a full lifetime.
Diabetes, PKU and childhood leukemia are not the absolute
death sentences they were in the past.
Should genetic screening for currently untreatable diseases
take the place of medical advances in removing deadly diseases from
the human race?
Further Investigations
Many scientific areas have applied molecular
biology tools to address questions.
Students who may not necessarily be interested in biology may
find one of the following applications more interesting.
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Forensic anthropologists attempt to identify the remains in
mass graves using a variety of methods including PCR.
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Epidemiologists studying new diseases and their sources will
employ molecular biology to analyze strains and variants.
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Penguins, and many other birds, must be sexed by DNA
analysis. Bird breeders, zoo keepers and veterinarians apply this
technology.
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Criminal investigators use PCR to amplify DNA samples from
crime scenes for further analysis.
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Plant and animal breeders wishing to verify the pedigree of
their stock use PCR and other molecular biology techniques.
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Lawyers specializing in legal issues arising regarding
intellectual property, privacy issues and appropriate use and
interpretation of evidence in criminal cases.
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Have students investigate genetic screening
in their families. Who has been screened and
what for? Infant
testing for PKU and other metabolic diseases has been in use for
about 40 years, so most of the students themselves have been
screened.
Have students determine expenses associated
with undiagnosed metabolic disorders including wages lost by
caregivers, medical expenses (personal and Medicare), insurance
costs and other costs such as special equipment or supplies required
to care for the patient. Compare
this cost with the cost of screening all infants.
Include the cost of the test and recommended treatment for
the patient.
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