Think about… 29.1 Recombinant DNA technology 29.2 D NA fingerprinting 29.3 Human Genome Project
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Transcript of Think about… 29.1 Recombinant DNA technology 29.2 D NA fingerprinting 29.3 Human Genome Project
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Victims of disasters can be identified by their fingerprints, dental information or birthmarks.
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These methods may not work when
the remains are incomplete.
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Identification may be done by matching the DNA.
DNA of the remains
reference DNA
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Identification may be done by matching the DNA.
DNA of the remains
reference DNA
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reference DNA
from personal items
from family members
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Why can DNA be used in identifyinga person1
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Why can DNA be obtained frompersonal items2
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Why can DNA samples from familymembers serve as reference in victim identification
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Think about…29.1 Recombinant DNA technology29.2 DNA fingerprinting29.3 Human Genome ProjectRecall ‘Think about…’Summary concept map
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29.1 Recombinant DNA technology
Applied genetics is about how the knowledge of genetics can be used for the good of society.
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29.1 Recombinant DNA technology
advances in biotechnology
(生物工程 )
recombinant DNA technology (重組 DNA 技
術 )
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29.1 Recombinant DNA technology
• techniques that a fragment of DNA from a donor cell or organism is isolated and inserted into the DNA of another cell or organism
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Domestication of plants and animals throughout human civilization
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29.1 Recombinant DNA technology
• can modify the characteristics of an organism more quickly and precisely
• allows transfer of new characteristics to organisms of different species
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29.1 Recombinant DNA technology
genetic engineering (遺傳工程 )
recombinant DNA technology (重組 DNA 技
術 )
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genetic engineering (遺傳工程 )• changing of the genetic
make-up of an organism by direct manipulation of genes or DNA
29.1 Recombinant DNA technology
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29.1 Recombinant DNA technology
How does recombinant DNA technology work?
There are three steps.
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29.1 Recombinant DNA technology
Obtain DNA fragments containing the gene of interest
donor cell
DNA containing gene of interest
gene of interest
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29.1 Recombinant DNA technology
Cut DNA fragments and plasmids with a specific enzyme
plasmid (質粒 )
• a small ring of extrachromosomal DNA in bacteria
• used as a vector (載體 )
bacterium
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29.1 Recombinant DNA technology
Cut DNA fragments and plasmids with a specific enzyme
gene of interest plasmid (質粒 )
enzymes
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29.1 Recombinant DNA technology
Join the gene of interest and plasmid together using an enzyme
gene of interest plasmid (質粒 )
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29.1 Recombinant DNA technology
Join the gene of interest and plasmid together using an enzyme
recombinant DNA (重組 DNA) / recombinant
plasmid (重組質粒 )
Animation
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29.1 Recombinant DNA technology
Join the gene of interest and plasmid together using an enzyme
introduce it into a host cell (宿主細胞 ) for replication and expression
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29.1 Recombinant DNA technology
Select the organisms that have acquired the gene of interest
Antibiotic resistance tests: The bacteria in the culture on the left are susceptible to the antibiotics contained in the white paper discs. The bacteria in the culture on the right are resistant to most of the antibiotics
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29.1 Recombinant DNA technology
What are the applications of recombinant DNA technology?Genetically modified organism (GMO) (基因改造生物 )• an organism whose genetic make-up
has been changed in a way that does not occur naturally by mating
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29.1 Recombinant DNA technology
What are the applications of recombinant DNA technology?Genetically modified food (GM food) (基因改造食物 )• food made from GMOs or their
components
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29.1 Recombinant DNA technology
1 Medicine• to produce pharmaceutical products
insulin production site
GM bacteria recombinant human insulin
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29.1 Recombinant DNA technology
• gene therapy (基因治療 )
vectors with a normal gene
cell with a defective gene
1 Medicine
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29.1 Recombinant DNA technology
2 Agriculture and food production
• to improve productivity and quality of farm animals and crops
non-GM salmon
GM salmon that grow faster
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29.1 Recombinant DNA technology
• to improve productivity and quality of farm animals and crops
GM soybean plants resistant to herbicides
2 Agriculture and food production
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29.1 Recombinant DNA technology
• to improve productivity and quality of farm animals and crops
GM tomatoes that have longer shelf life
2 Agriculture and food production
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29.1 Recombinant DNA technology
• to improve productivity and quality of farm animals and crops
GM rice that has more vitamin
2 Agriculture and food production
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29.1 Recombinant DNA technology
3 Chemical industries• to produce GM microorganisms that can
produce the following more efficiently:
enzymes
amino acids
polysaccharides
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29.1 Recombinant DNA technology
4 Environmental protection• to produce GM bacteria that can break
down oil spills more efficiently
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29.1 Recombinant DNA technology
5 Ore mining• to produce GM bacteria that can extract
metals more efficiently from ores• Eg.bacteria catalyse the breakdown of the mineral pyrite (FeS2) by oxidising
the sulfur and metal (in this case ferrous iron, (Fe2+) using oxygen.
• This yields soluble products that can be further purified and refined to yield the desired metal.
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1 is the DNA that results from the combination of DNA fragments from two different cells or organisms.
Recombinant DNA
29.1 Recombinant DNA technology
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2 refers to the techniques that a fragment of DNA from a donor cell or organism is isolated and inserted into the DNA of another cell or organism.
Recombinant DNA technology
29.1 Recombinant DNA technology
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a Obtain DNA fragments containing the from donor cells through proper processes.
3 Major steps in recombinant DNA technology:
gene of interest
29.1 Recombinant DNA technology
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b Cut the DNA containing the gene of interest with a specific
. Cut open a plasmid using the same .enzyme
29.1 Recombinant DNA technology
4 Major steps in recombinant DNA technology:
enzyme
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c Join the and the together with the help of another enzyme.
D _____ the organisms that acquire the gene of interest
gene of interestplasmid
29.1 Recombinant DNA technology
4 Major steps in recombinant DNA technology:
select
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a To produce products and for
in medicine.
4 Applications of recombinant DNA technology:
gene therapy
29.1 Recombinant DNA technology
pharmaceutical
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b To improve the and of farm animals and
crops in agriculture.
productivityquality
29.1 Recombinant DNA technology
4 Applications of recombinant DNA technology:
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c To produce in food production. food
genetically modified
29.1 Recombinant DNA technology
4 Applications of recombinant DNA technology:
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d To produce that can
produce enzymes, amino acids or polysaccharides more efficiently.
organismsgenetically modified
29.1 Recombinant DNA technology
4 Applications of recombinant DNA technology:
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e To produce genetically modified organisms that can break down
more efficiently.oil spills
29.1 Recombinant DNA technology
4 Applications of recombinant DNA technology:
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f To produce genetically modified organisms that can extract more efficiently from ores.
metals
29.1 Recombinant DNA technology
4 Applications of recombinant DNA technology:
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29.2 DNA fingerprinting• techniques involved in the use of
DNA analyses to identify individuals
How do scientists produce DNA fingerprints?
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29.2 DNA fingerprinting
Extract DNA
DNA
cells in blood
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29.2 DNA fingerprinting
Obtain DNA fragments containing the highly variable regions
size varies among individuals
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29.2 DNA fingerprinting
Separate DNA fragments according to their size
gel electrophoresis (凝膠電泳 )
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29.2 DNA fingerprinting
Produce DNA fingerprints
DNA bands• pattern is unique to
each individual (except identical twins)
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• uses an electric field to drive DNA fragments to the positive terminal
29.2 DNA fingerprinting
Gel electrophoresis
negatively charged!
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• molecular space in the gel allows shorter DNA fragments to move faster than the longer ones
29.2 DNA fingerprinting
Gel electrophoresis
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• molecular space in the gel allows shorter DNA fragments to move faster than the longer ones
29.2 DNA fingerprinting
Gel electrophoresis
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• molecular space in the gel allows shorter DNA fragments to move faster than the longer ones
29.2 DNA fingerprinting
Gel electrophoresis
longer fragments
shorter fragments
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• DNA fragments separated into bands according to their size
29.2 DNA fingerprinting
Gel electrophoresis
longer fragments
shorter fragments
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• staining is required before the DNA bands can been seen
29.2 DNA fingerprinting
Gel electrophoresis
fluorescent under UV light
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Mix 80 cm3 of 5X TBE buffer solution and 320 cm3 of distilled water in a beaker.
29.1Separation of DNA fragments using gel electrophoresisA Preparation of 1X TBE buffer solution
Video
29.2 DNA fingerprinting
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29.1
1 Mix 60 cm3 of 1X TBE buffer solution and 0.9 g of agarose powder in a conical flask.
B Preparation of agarose gel
2 Heat the mixture over a hotplate until the agarose powder completely dissolves.
29.2 DNA fingerprinting
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29.1
3 Cool down the agarose solution to about 60ºC.
4 Seal the two ends of the gel mould with sticky tape and pour the agarose solution into the gel mould slowly.
29.2 DNA fingerprinting
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29.1
5 Insert the gel comb into the gel mould at one side.
6 After about 30 minutes, remove the gel comb and sticky tape carefully.
29.2 DNA fingerprinting
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29.1
1 Label the six tubes of DNA samples A to F.
C Gel electrophoresis of DNA fragments
2 Put the gel with the gel mould in the electrophoresis tank. The side with the wells should be placed at the cathode (-) of the tank.
29.2 DNA fingerprinting
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29.1
3 Add 1X TBE buffer solution until the gel is covered.
29.2 DNA fingerprinting
4 Starting from the leftmost well, slowly load the DNA marker, DNA samples A to F into separate wells using a syringe fitted with a plastic tip. Wash the syringe with a new buffer solution between successive loadings.
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29.1
5 Cover the lid of the electrophoresis tank. Connect the electrodes and turn on the power supply to apply a 100 V d.c. voltage across the gel.
29.2 DNA fingerprinting
100 V
- +
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29.1
6 After about 60 minutes, when the tracking dyes reach about half to two thirds of the length of the gel, turn off the power supply.
29.2 DNA fingerprinting
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29.1
7 Put the gel into a plastic lunch box containing methylene blue solution. Leave the gel for about 12 hours.
29.2 DNA fingerprinting
8 Take a photo of the gel with DNA bandings.
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1 Forensic science (法證科學 )
29.2 DNA fingerprinting
Applications of DNA fingerprinting
• to provide evidence to the court
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2 Victim identification
29.2 DNA fingerprinting
Applications of DNA fingerprinting
• to identify victims in disasters
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3 Parentage test (親子鑑證 )
29.2 DNA fingerprinting
Applications of DNA fingerprinting
• to establish family relationships
Animation
father mother child
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1 refers to the techniques involved in the use of DNA analyses to identify individuals.
DNA fingerprinting
29.2 DNA fingerprinting
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2 Steps in DNA fingerprinting:
a Extract from samples.
b Obtain containing highly variable
regions through proper processes.
DNA
DNA fragments
29.2 DNA fingerprinting
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c Separate DNA fragments using .
d Produce .
gel electrophoresisDNA fingerprints
29.2 DNA fingerprinting
2 Steps in DNA fingerprinting:
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3 Gel electrophoresis makes use of an electric field to drive DNA fragments to the terminal. DNA fragments move at speeds that depend on their .
positive size
29.2 DNA fingerprinting
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4 Applications of DNA fingerprinting:
a To provide evidence to the court in .
b To in disasters. c To establish family
relationships in .
forensic scienceidentify victims
parentage tests
29.2 DNA fingerprinting
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• genome (基因組 ):- DNA sequence on one set of
chromosomes in an organism
29.3 Human Genome Project
•A colorized electron micrograph of Haemophilus influenzae, a bacterium that was the first free-living organism to have its genome sequenced, in 1995.•This bacterium causes respiratory infections and bacterial meningitis (inflammation of the protective membranes of the brain) in humans.
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• research project started in 1990What is the HGP about?• involved scientists from >18 countries
29.3 Human Genome Project
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• goals:- DNA sequencing of human DNA- mapping of human gene loci- develop new technologies e.g
diagnostic or therapeutic techniques
- address the ethical (道德倫理的 ), legal and social issues
29.3 Human Genome Project
What is the HGP about?
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29.3 Human Genome Project
What is the HGP about?
HGP started
1990
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29.3 Human Genome Project
What is the HGP about?
sequencing of yeast genome and mouse genome completed
1996
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29.3 Human Genome Project
What is the HGP about?
sequencing of first human chromosome completed
1999
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29.3 Human Genome Project
What is the HGP about?
draft version of human genome sequence completed
2000
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29.3 Human Genome Project
What is the HGP about?
finished version of human genome sequence completed
2003
• all goals achieved, two years earlier than expected!
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29.3 Human Genome Project
What is the HGP about?
2003
• all goals achieved, two years earlier than expected!
analyses of data continue
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The human genome project
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1 Better understanding of geneticsBenefits of the HGP
2 Improved diagnoses and treatment of diseases
29.3 Human Genome Project
breast cancer
gene related to
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3 Better understanding of evolutionBenefits of the HGP
29.3 Human Genome Project
similarity in base sequence
98% 85%
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3 Better understanding of evolutionBenefits of the HGP
29.3 Human Genome Project
more closely related
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1 The genetic data obtained may still not be enough to understand all biological processes.
Limitations of the HGP
2 It has raised ethical, legal and social issues.
The Ethical, Legal, and Social Implications (ELSI) program as an integral part of the Human Genome Project.
29.3 Human Genome Project
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Example:Limitations of the HGP
29.3 Human Genome Project
Who is the owner of personal genetic information?
I worked it out.
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Example:Limitations of the HGP
29.3 Human Genome Project
Who is the owner of personal genetic information?
I developed the technology.
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Example:Limitations of the HGP
29.3 Human Genome Project
Who is the owner of personal genetic information?
It’s my personal information.
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Example:Limitations of the HGP
29.3 Human Genome Project
Who can gain access to personal genetic information?
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Example:Limitations of the HGP
29.3 Human Genome Project
Would the diagnosis of an incurable disease cause anxiety?
I’m sorry. You have the faulty gene and there is no cure for the disease.
Oh no.
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Was that a rational decision?•After discovering a mutated BRCA1 gene, known to increase the likelihood of breast cancer 60 to 80 percent, actress Angelina Jolie underwent a radical preventive double mastectomy.
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Example:Limitations of the HGP
29.3 Human Genome Project
How does the personal genetic information affect the public perception of an individual?
Go! He has a gene related to violence.
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a and of the human genome.
b Developing related new .
Sequencing
technologies
mapping
29.3 Human Genome Project
1 Goals of the Human Genome Project (HGP):
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29.3 Human Genome Project
1 Goals of the Human Genome Project (HGP):
c Addressing the , and issues.
ethicalsocial
legal
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29.3 Human Genome Project
2 The contributions of the data obtained from the HGP:
a Better understanding of
. b Improved and of diseases.
genetics
treatmentdiagnoses
c Better understanding of . evolution
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still not be enough to understand all .
29.3 Human Genome Project
3 The limitations of the data obtained from the HGP:
a The genetic data obtained may
biological processesb They have raised ethical,
legal and social issues.
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Why can DNA be used in identifyinga person?1
Like fingerprints, the DNA sequence is unique to each individual (except identical twins).
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Why can DNA be obtained frompersonal items?2Samples such as blood and hair with hair roots may be left on such personal items.
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Why can DNA samples from familymembers serve as reference in victim identification?
3The closer the family relationship, the more similar are the DNA sequences.
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is aboutApplied genetics
applications of genetics knowledge
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applied in
recombinant DNA technology
examples include
applications of genetics knowledge
production of GMOs
pharmaceutical products
GM food
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applied inDNA fingerprinting
applications of genetics knowledge
forensic science
victim identification
parentage test
examples include
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raisesHuman Genome Project
applications of genetics knowledge
ethical, legal and social issues
examples include
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better understanding
of genetics
better understanding
of evolution
improved diagnoses and treatment of diseases
data contributes to Human Genome Project