dna kit instructions
TRANSCRIPT
8/6/2019 DNA Kit Instructions
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DNA is the longest molecule knownand it exists in every living cell. Thecode it carries determines the form
and behavior of all life on earth.
When released from a cell, DNAtypically breaks up into filaments.
In solution, these strands have a
slight negative electric charge,
which makes for some fascinating
chemistry. For example, the more
negative sections of one DNA
strand will tend to attract the more
positive regions of another. This
causes these DNA floatingmolecules to clump together into a
big gooey mass that you can easilysee. However, if salt is added, the
salt ions are attracted to thenegative charges on DNA,
effectively neutralizing them. This
stops the separate fragments from
sticking together and keeps them
floating about in solution.
So by controlling the salt
concentration, biologists can make
Your Kit Contains…For the buffer solution
• Powdered Buffer: Contains 1)
DNA Suspension Agent —
21.5% NaCl (pure table salt) to
lubricate the DNA molecules to
keep them from sticking
together. 2) pH Stabilizer:
77.3% NaHCO3 (purified baking
soda) to neutralize any acids in
the organic matter. 3) Protein
Destroyer: 1.2% Papainenzyme, eats proteins so theydon't contaminate your DNA.
• Cell Blaster: sodium dodecylsulfate—a detergent that rips
up cell walls and nuclei so the
molecules inside can seep out.
To Stain the DNA
Blue Monster DNA stain to make
the DNA clearly visible.
Extract DNA Experimenter Kit Age 12+
DNA fragments either disperse or
glom together. And therein lies the
secret of separating DNA from cells.
The materials I've provided you
in my Super DNA Experimenter's
Kit will make it easy for you to
extract and purify in your kitchenDNA from living things exactly as
professional scientists do in their
laboratories. Pretty cool, eh?
Sundries
Graduated Test Tube to harvest
the DNA and measure your yield.
Nylon Fast-flow Filter to remove
the gunk after you've extracted the
organic molecules from the cells.
Glass Extraction Rod to remove
the DNA from the test tube.
Plastic 4 oz. squeeze bottle to
conveniently hold and dispenserubbing alcohol.
Three 250 ml Tri-pour Beakers to mix and process the materials.
Not included
Rubbing or Isopropyl Alcohol:
Isopropyl alcohol is a “hazardous” material and is far too expensive to
ship. Use the highest concentrationthat your drugstore sells. NOTE:
You MUST chill your alcohol in thefreezer before you begin!
Did you know…
DNA stands for
Deoxyribonucleic
Acid
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Dr. Shawn's Experimenter's Kits: Extract DNA in Your Kitchen
While far more challenging thanexperiments with quantity, you can
also experiment with DNA itself.You'll first need to remove the DNA
sludge from the original buffersolution that is full of organic
contaminants and re-dissolve it in a
batch of fresh buffer. Moreover, if
you can't complete your experiment
in one sitting you'll want to store
your DNA for later. Also, it's often
useful to stain your DNA to make it
easier to see. Here's how to do all
of these important steps.
How to Remove the DNA
First, clean and dry the glassextraction rod thoroughly making
certain it is completely free of oils
or dirt or chemical contaminants of
any kind. You should soap, rub and
carefully rinse it in distilled or
bottled water and dry it completely
with a fresh paper towel.
Next, rest the test tube inside aglass so you'll have two hands free.
Then gently insert the clean rod
through the DNA gunk with its tip just below the boundary of the
buffer solution. Hold the rod stillwith one hand and with the otherone very slowly twirl the rod round
and round in the same direction.
Longer pieces of DNA will spool
onto the rod just like spaghetti on a
fork, leaving smaller fragments
Experiments With DNA
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behind. After a minute or so, pull
the rod slowly up through the
alcohol. The alcohol will make theDNA adhere to the glass. When you
get it into the air you will see a
transparent viscous "snot-like"
sludge clinging to the rod.There is a bit of artistry required
here so don't get frustrated if it
doesn't work at first. In fact,
usually only a small fraction of yourDNA will adhere to the rod. The
picture below shows a typical result
for a novice molecular biologist, sodon't get discouraged. Your results
will improve with practice and
experience.
Re-dissolving DNA You can re-dissolve your DNA in
fresh buffer to continue your
experiments. Since you aren't
extracting DNA from cells at this
stage, the fresh buffer need only
contain the Suspension Agent and the pH Stabilizer. And remember,
less is more here; the less bufferyou use the more of the re-
suspended DNA you will be able to
extract later.
Store Your DNA For Later It’s easy to store your DNA.
Just place the viscous sludge that
you twirled up on your stick in a
container filled with ice-cold
isopropyl (rubbing) alcohol, then
lace the container in the freezer.
Use rubbing alcohol
isopropyl from your local drstore. When stored in chill
alcohol like this, your DNA w
keep almost forever.
Dyeing DNA Even after the mo
thorough extraction, som
residual DNA will linger in t
vessel, forming an invisib
cobweb within the liquid. B
with a little more effort, ycan see that material, too.
Dr. Shawn's special Bl
Monster DNA Stain, which included in yo
Experimenter's Kit, bin
directly to charged Dfragments. A tiny amou
added to the remaini
solution will thus stain tendr
of uncollected DNA. Add only
tiny droplet: you want all t
dye molecules to bind to t
DNA, with none left over
stain the water. If you hatrouble measuring the quant
of DNA you’ve extracted,
ahead and drop a droplet the Blue Monster Stain in
the graduated cylinder aftyou've precipitated the DN
The droplet will fall straig
through the alcohol and cli
beautifully to the DNA.
The blue band marks the layer of
DNA sludge when stained.
DNA extracted rom a cow's brain is visible on the end o the Extraction Rod.
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Remember—Learning follows
interest. That means that it is
hard to learn very much or do
very well with projects that
don't interest you. So make
sure to pick a project that
really gets your juices flowing!
Doing a science fair project? Then you'll need a question to answer
or hypothesis to pose. Here are some ideas for proven championshipscience fair projects. If none of these appeal to you, that's fine. Learn
more and ask your own questions. Doing what interests you is the
always the best way to approach any science project.
Note: Since most experimenters find it hard to get the DNA out of the test tube, “Type One” projects are much easier than “Type Two.” So
if you want high certainty of success, stick with Type One.
Ideas for Science Projects
Procedure: Extract DNA from different varieties
of fruits, vegetables, fungi and other material in
your kitchen, garden, butcher shop or the greatoutdoors. Pick at least three different sources.
In addition to fruits and vegetables, try flour,
egg whites or egg yokes, hamburger, liver,outdoor plants, and so on.
Extract DNA from equal amounts of each one
and carefully compare the quantities you find.
Questions to consider…
1) What kinds of plants yield the most DNA
per unit volume: Fruits, vegetables, orlegumes?
2) What part of a plant produces the most
DNA per unit volume: Flowers, stems,
fruits, nuts or leaves?3) How does temperature affect DNA in vivo
(in life)? Use a cooking thermometer to setthe temp of water on a stove. Soak
Type One: Comparing Amounts of DNA Produced Under Different Conditions
Type Two: Experiments With DNA Itself
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Procedure: Here's what you need to do.a) First, extract a large amount of DNA from
onions, bananas or another copious source,
and store it in isopropyl alcohol in your
freezer.
b) When you're ready to do your experiment,
redissolve a quantity of DNA, say 5 ml (1tsp), into each of two beakers containing
identical amounts of fresh buffer.c) You must treat both samples exactly the
same except for just one thing. You expose
one sample, your "test" sample, tosomething that you think might destroy
DNA. The other, your "control" sample, you
leave alone. Run all samples in pairs side
by side. Every test sample must have its
own control.
whatever you’re experimenting with in the
bath for several minutes. Remove and chill
with ice water. Then find out how muchDNA you can extract. Do this at several
temps between room temperature and
boiling and graph your results.4) Do organic vegetables yield the same
amount of DNA as non-organic?
5) Do fruits yield more extractable DNA when
they are underripe, ripe or overripe?
6) Does the fraction of DNA change for
different types of fungi, or do all fungi
yield about the same amount per unit
volume?7) Try changing the procedure. How much
DNA do you get when you use tap water,
or use different amounts of detergent, or a
different detergent, etc.?Not interested in any of these questions? Then
see if they spark any ideas of your own that do
interest you!
d) Finally, reconstitute the DNA in bothsamples by adding isopropyl alcohol.
Compare the amount recovered from thetest to the control sample to discover how
much DNA, if any, was destroyed during
your trial.
Questions to consider…
1) How rapidly does DNA degrade when
exposed to sunlight in vitro (in a test tube)?
Expose different test samples for different
amounts of time to outdoor sunlight. Thin
plastic containers let ultraviolet through.2) How rapidly does DNA degrade at different
temperatures in vitro? At a giventemperature, plot fraction lost vs. time.
3) How do chemicals affect DNA in vitro? Plot
the fraction of DNA recovered when
exposed to small concentrations of chlorine
(bleach) and other household chemicals.
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Need More Info about DNA? There's tons of great information about
DNA online. To find what's current, search "DNA education" on theInternet.
Resources…
Contact us:
Bright Science, LLC
1356 Saxon Lane
Naperville, IL 60564
Phone: 630-300-3966
Email:
We’re on the Web!
For help with any science
project :
www.scifair.org
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This material is copyrighted
by Dr. Shawn (Shawn
Carlson, Ph.D.). All rights are
reserved.