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Biology
Module 6
Manipulating Genomes
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emma
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Cards (33)
What is PCR used for?
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 for PCR?
DNA sample
Free DNA nucleotides
Primers
DNA polymerase from
thermophilic bacteria
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At what temperature does DNA strands separate 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 in PCR?
72°C
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How does DNA polymerase function during 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 purpose of adding a fluorescent molecule to DNA?
To visualize DNA under
UV light
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How does gel electrophoresis separate DNA fragments?
By length, due to DNA's
negative charge
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What are the steps 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?
Sequences that read the same
forwards
and
backwards
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What are the two ways restriction enzymes can cut DNA?
Blunt ends
or
sticky ends
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How do scientists extract a gene of interest using restriction enzymes?
By analyzing
recognition sites
around the gene
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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 determine besides criminal identification?
Genetic
relationships between individuals
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What is the process of making insulin using genetic engineering?
Remove insulin gene from human DNA
Cut plasmid with
restriction enzymes
Join sticky ends with
DNA ligase
Mix
recombinant plasmid
with bacteria
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
synthesizes
the drug
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How are genetically modified animals produced?
Inject gene into
fertilized egg nucleus
Implant into
adult animal
All
cells develop
with
drug gene
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What are the arguments for using GMOs?
Increase
crop yield
and nutrition
Reduce
pesticide
use
Produce human proteins without
allergies
Stable vaccines for remote areas
Cheaper
industrial enzymes
Cost-effective
drug production
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What are the arguments against using GMOs?
Patented
seeds
are 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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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 some problems with gene therapy?
Immune response
,
insertion errors
, and cost
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What is the chain-termination method used for?
To sequence
DNA
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How does the chain-termination method work?
Mix DNA sample, primers,
polymerase
,
nucleotides
Add fluorescently-labelled modified nucleotides
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)
Amplify
BACs in bacteria
Purify and sequence fragments
Use
software
to reconstruct genome
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What is synthetic biology?
Building proteins from
DNA sequences
Example: Synthetic artemisinin from yeast
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What are the applications of comparing DNA sequences?
Epidemiology
: Identify disease-linked
genes
Evolutionary relationships: Assess
organism
relatedness
Genotype-phenotype
relationships: Predict
traits
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