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Biology Module 6
Manipulating genomes
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Cards (34)
What is the purpose of PCR?
To
amplify
fragments
of
DNA
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Why is PCR important for crime scene analysis?
It
amplifies
insufficient
DNA samples
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What components are needed to prepare a PCR mixture?
DNA
sample
Free DNA nucleotides
Primers
DNA polymerase from thermophilic bacteria
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At what temperature does DNA separation occur in PCR?
95°C
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What happens during the annealing stage of PCR?
The
primer
binds to the DNA
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What is the optimum temperature for DNA polymerase during PCR?
72°C
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How does DNA polymerase contribute to DNA synthesis in PCR?
It catalyzes
phosphodiester
bond formation
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How many cycles of PCR are typically carried out?
30-40
cycles
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What is the role of ethidium bromide in electrophoresis?
It binds to
DNA
and makes it visible
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Why does DNA move towards the positive charge in gel electrophoresis?
DNA is
negatively
charged
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How do shorter DNA fragments behave in gel electrophoresis?
They travel further than
larger
fragments
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What are the steps involved in gel electrophoresis?
Prepare
agarose gel
with wells
Place gel in
buffer solution
Mix DNA with
loading dye
Pipette DNA into wells
Apply
electrical current
Visualize banding pattern under
UV light
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What do restriction enzymes do?
Cut
double-stranded
DNA at
specific sequences
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What are palindromic recognition sites in DNA?
They read the same
forwards
and
backwards
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What are the two ways restriction enzymes can cut DNA?
Creating
blunt ends
or
sticky ends
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How do scientists use restriction enzymes in genetic manipulation?
To extract
genes
from
longer DNA sections
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What is DNA profiling used for?
To
analyze
and
compare
DNA samples
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What is the first step in DNA profiling?
Collecting
a
DNA
sample
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How are PCR products separated in DNA profiling?
Using
gel electrophoresis
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What can DNA profiling reveal about individuals?
Genetic
relationships and
evolutionary
links
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What is the process of genetic engineering for insulin production?
Remove insulin gene from human DNA
Cut plasmid with
restriction enzymes
Join sticky ends with
DNA ligase
Mix
recombinant
plasmid with bacteria
Use
electroporator
to increase permeability
Grow
transgenic
bacteria in fermenters
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How are plants genetically modified to produce drugs?
Create
GM
bacterium with
drug gene
Bacterium
infects
plant cell
Inserts DNA into
plant genome
Plant grows with drug-producing gene
Drug can be
extracted
or consumed
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How are genetically modified animals produced?
Inject gene into fertilized
egg cell
Implant into
adult animal
All cells develop with
drug-producing
gene
Protein
is purified from
milk
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What are the arguments for and against GMOs?
Arguments for:
Increased
crop yield
and nutrition
Pest-resistance reduces
pesticide
use
Human
proteins
reduce allergic reactions
Vaccines stable without refrigeration
Cheaper
industrial processes
Arguments against:
Patents
make seeds expensive
Risk of creating
'superweeds'
Ethical concerns about
manipulation
Religious objections to 'playing God'
Unforeseen
long-term
impacts
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What is gene therapy used for?
To treat
genetic
diseases
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How is gene therapy applied for recessive diseases?
A
functional gene copy
is introduced
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What is the difference between germ line and somatic gene therapy?
Germ line affects
gametes
; somatic affects
body
cells
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What are the problems associated with somatic gene therapy?
Requires multiple
insertions
and may trigger
immune response
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What is the chain-termination method used for?
To sequence
DNA
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What are the steps in the chain-termination method for DNA sequencing?
Mix DNA sample, primers, polymerase, nucleotides
Add
modified nucleotides
with fluorescent tags
Carry out
PCR
to create varying lengths
Use
electrophoresis
to separate fragments
Read sequence from gel
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What is the purpose of high-throughput sequencing?
To sequence
DNA
quickly and cheaply
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What is whole genome sequencing?
Fragment DNA using
restriction enzymes
Insert fragments into bacterial
artificial chromosomes
(
BACs
)
Insert BACs into bacteria for replication
Purify and further fragment DNA
Sequence shorter fragments and
assemble genome
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What is synthetic biology?
Use
DNA sequences
to build
proteins
Example:
Synthetic artemisinin
from
yeast
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What are the applications of comparing DNA sequences?
Epidemiology: Identify disease-linked
genes
Evolutionary relationships: Determine
organism
closeness
Genotype-phenotype relationships
: Predict traits from genotype
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