Download Teacher quality grant

Biology Partnership
(A Teacher Quality Grant)
Biotechnology
Hands-on Investigations
Nancy Dow
Jill Hansen
Tammy Stundon
May 11, 2013
Gulf Coast State College
Panhandle Area Educational Consortium
5230 West Highway 98
753 West Boulevard
Panama City, Florida 32401
Chipley, Florida 32428
850-769-1551
877-873-7232
www.gulfcoast.edu
Pre-test????
Q and A board
What is Biotechnology?
How do we use Biotechnology?
Florida Next Generation
Sunshine State Standards
SC.912.L.16.10 Evaluate the impact of biotechnology on the
individual, society and the environment, including medical and
ethical issues.
Clarification
• Students will evaluate examples and/or explain the possible impact
of biotechnology on the individual, society, and/or the
environment.
Content Limits
• Items may assess current issues but will not require knowledge of
specific biotechnologies or specific medical issues.
• Items assessing the possible impacts of biotechnology will not
assess monetary impacts.
Crossing -over
- Occurs during Prophase I of
Meiosis
- Between chromatids of
homologous pairs
- Increase genetic variation
DNA
- Function
- Components
- Location
U.S. Department of Energy Genomics:GTL Program
http://www.ornl.gov/hgmis
Biotechnology
• Genetic engineering
– def: use of technology to alter the
genes of viruses, bacteria, and other
cells for medical or industrial
purposes to better the quality of life
– Altering genes of unicellular
organisms and plants and animals
• Genome: composed of DNA, is our hereditary
code (the “blueprint”)
• Molecular biology: the study of genes and the
molecular details that regulate the flow of
genetic information from DNA to RNA to
proteins, from generation to generation.
Biotechnology uses this knowledge to manipulate
organisms’ DNA to help solve human problems.
Caught by a Kiss!
Forensic Science!
Ready to use what you know in the ‘real world’?
Gel Electrophoresis
• Technique used to
separate DNA bands
based on size
• Comparison of sample
bands to markers
allows…
– Visible confirmation of
desired product
– Quantification of
sample DNA
How does it work?
•
Electrolysis: the
splitting of water using
electricity
•
Electrophoresis: a
method of separating
charged molecules in
an electrical field;
DNA has an overall
negative charge
Used to separate DNA
fragments by size
Why Run a Gel?
DNA fingerprints are useful
in several areas of society.
They are used by
professionals in human
health and the justice
system.
1. Diagnosis of
inherited disorders
2. Developing cures for
inherited disorders
3. Forensic or criminal
4. Personal identification
1.
2.
3.
4.
Extract DNA,
suspend in buffer
Restriction
enzymes search
and isolate unique
sequences found in
your genome.
Everyone has a
different number of
these sequences
found at different
locations
PCR replicates
your unique
sequences so we
have enough to
visualize
Result is several
different pieces of
DNA of differing
How do we visualize DNA?
Steps of Gel Electrophoresis
Prepare agarose gel
(just like jello – but not as tasty)
Pour into casting tray with comb and allow to solidify
Add running buffer, load samples and marker
Run gel at constant voltage until band separation occurs
View DNA on UV light box and document results
Electrophoresis Equipment
Power supply
Gel tank
Cover
Electrical leads

Casting tray
Gel combs
Agarose Gel
• A porous material derived from red
seaweed
• Acts as a sieve for separating DNA
fragments; smaller fragments travel
faster than large fragments
1% agarose
• Concentration affects DNA migration
– Low conc. = larger pores better
resolution of larger DNA fragments
– High conc. = smaller pores better
resolution of smaller DNA fragments
2% agarose
Loading Dye
• DNA samples are loaded into
a gel after the tank has been
filled with buffer, covering
the gel
• Contains a dense substance,
such as glycerol, to allow the
sample to "fall" into the
sample wells
• Contains one or two tracking
dyes, which migrate in the
gel and allow monitoring of
how far the electrophoresis
has proceeded.
Who Dunnit?
Virtual Gel - Guided Inquiry
POLYMERASE CHAIN REACTION
(PCR)
- 1983
- With PCR, you can target and make millions of
copies (amplify) a specific piece of DNA (or gene)
out of a complete genome.
- PCR impacted several areas of genetic research:
- as a medical diagnostic tool to detect specific mutations that may cause
genetic disease
- in criminal investigations and courts of law to identify suspects
- in the sequencing of the human genome
PCR Amplification
• PCR makes use of the same
basic processes that cells use
to duplicate their DNA
(replication)
– Complementary DNA strand
hybridization
– DNA strand synthesis via DNA
polymerase
PCR intro
Recipe for PCR Amplification
1. DNA sample: containing the intact sequence of DNA to be
amplified
2. Master mix:
√ Free nucleotides (dNTPs): raw material of DNA (A,T,C,G)
√ DNA
polymerase (Taq polymerase): enzyme that assembles
the nucleotides into a new DNA chain
√ Primers:
pieces of DNA complementary to the template that
tell DNA polymerase exactly where to start
√ Fluorescent dye: “lights up” when it binds to complete DNA
strands
3. Thermocycler: used to rapidly heat and cool
DNA samples to facilitate DNA amplification.
Steps of
PCR
1. Temperature separates
the strand
↑ temperature
2. Primers attach like
Bookends
↓ temperature
3. Taq takes free
nucleotides & adds them
to the ends of the primer
↑ temperature
Step 3 is also called Elongation
Results of
PCR
With 30 cycles the
DNA is amplified
exponentially
Results in 1.1x1012
sets of preciselength DNA
PCR Song
Manipulating the DNA
Bt CORN
Bacterium (Bacillus thuringiensis)
Bt crystal gene
Plant cell
1
Corn plant destroyed
by insect pests
Corn genome
2 Bacterial gene coding for Bt
crystals, which are poisonous to
the insect pests, is inserted
directly into the corn plant’s DNA.
3 Bt crystals—toxic to insect pests—
are now produced by the corn plant
itself, reducing the amount of
pesticides the farmer must use.
Manipulating
the DNA
Almost 10% of the
world’s people suffer
from vitamin A
deficiencies—leading
to 250,00 cases of
blindness each year.
The addition of betacarotene-producing
genes to white rice
has increased its
vitamin A content
almost 25-fold.
GOLDEN RICE
Genes in the pathway for
beta-carotene production
are introduced into the
white rice genome.
Daffodil
Bacteria
White rice
Beta-carotene
Golden rice
Manipulating the DNA
GENETICALLY MODIFIED CROPS IN THE UNITED STATES
Corn
Cotton
55%
45%
Soybeans
24%
76%
15%
85%
Proportion of crops that are not genetically modified
Proportion of crops that are genetically modified
Commonly called GM Foods
• GMO Computer lab
“
– Selective breeding
– Transgenic manipulations
– Guess what’s coming to dinner
GMO Debate
Your scientists were so preoccupied with whether or not they
could, they didn't stop to think if they should.”
Dr. Ian Malcolm, Jurassic Park
• Should we or shouldn’t we……
Jurassic Park Mr. DNA clip
Follow up
•Q & A
•Post Test
•
•
•
DNA Science: by David A.
Micklos and Greg A. Freyer
Walkthrough with Quiz
Virtual Lab