dna kit instructions

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



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.



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.



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:

[email protected]

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.

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