You just need a drop or two, because a little bit of enzyme will go a

Trouble-shooting long way. Enzymes are fast and powerful!

1. I don't think I'm seeing DNA. What should I be looking for? 8. Why does the DNA clump together?

Look closely. Your DNA may be lingering between the two layers DNA precipitates when in the presence of alcohol, which means it
of alcohol and pea soup. Try to help the DNA rise to the top, doesn't dissolve in alcohol. This causes the DNA to clump together
alcohol layer. Dip a wooden stick into the pea soup and slowly pull when there is a lot of it. And, usually, cells contain a lot of it!
upward into the alcohol layer. Also, look very closely at the
alcohol layer for tiny bubbles. Even if your yield of DNA is low, For example, each cell in the human body contains 46
clumps of DNA may be loosely attached to the bubbles. chromosomes (or 46 DNA molecules). If you lined up those DNA
molecules end to end, a single cell would contain six feet of DNA!
2. What can I do to increase my yield of DNA? If the human body is made of about 100 trillion cells, each of
which contains six feet of DNA, our bodies contain more than a
Allow more time for each step to complete. Make sure to let the billion miles of DNA!
detergent sit for at least five minutes. If the cell and nuclear
membranes are still intact, the DNA will be stuck in the bottom 9. How can we confirm the white, stringy stuff is DNA?
layer. Or, try letting the test tube of pea mixture and alcohol sit for
30-60 minutes. You may see more DNA precipitate into the There is a protocol that would allow you to stain nucleic acids, but
alcohol layer over time. the chemical used would need to be handled by a teacher or an
adult. So, for now, you'll just have to trust that the molecules
Keep it cold. Using ice-cold water and ice-cold alcohol will precipitating in the alcohol are nucleic acids.
increase your yield of DNA. The cold water protects the DNA by
slowing down enzymes that can break it apart. The cold alcohol 10. Isn't the white, stringy stuff actually a mix of DNA and
helps the DNA precipitate (solidify and appear) more quickly. RNA?

Make sure that you started with enough DNA. Many food That's exactly right! The procedure for DNA extraction is really a
sources of DNA, such as grapes, also contain a lot of water. If the procedure for nucleic acid extraction.
blended cell soup is too watery, there won't be enough DNA to see.
To fix this, go back to the first step and add less water. The cell 11. How long will my DNA last? Will it eventually degrade and
soup should be opaque, meaning that you can't see through it. disappear?
Understanding the Science behind Your DNA may last for years if you store it in alcohol in a tightly-
sealed container. If it is shaken, the DNA strands will break into
the Protocol smaller pieces, making the DNA harder to see. If it disappears it's
likely because enzymes are still present that are breaking apart the
3. Why add salt? What is its purpose? DNA in your sample.

Salty water helps the DNA precipitate (solidify and appear) when Using more sophisticated chemicals in a lab, it is possible to obtain
alcohol is added. a sample of DNA that is very pure. DNA purified in this way is
actually quite stable and will remain intact for months or years.
4. Why is cold water better than warm water for extracting
DNA? Comparing the DNA Extracted from
Cold water helps keep the DNA intact during the extraction
process. How? Cooling slows down enzymatic reactions. This
Different Cell Types
protects DNA from enzymes that can destroy it. 12. Does chromosome number noticeably affect the mass of
DNA you'll see?
Why would a cell contain enzymes that destroy DNA? These
enzymes are present in the cell cytoplasm (not the nucleus) to Cells with more chromosomes contain relatively more DNA, but
destroy the DNA of viruses that may enter our cells and make us the difference will not likely be noticeable to the eye. The amount
sick. A cell's DNA is usually protected from such enzymes (called of DNA you will see depends more on the ratio of DNA to cell
DNases) by the nuclear membrane, but adding detergent destroys volume.
that membrane.
For example, plant seeds yield a lot of DNA because they have
5. How is the cell wall of plant cells broken down? very little water in the cell cytoplasm. That is, they have a small
volume. So the DNA is relatively concentrated. You don't have to
It is broken down by the motion and physical force of the blender. use very many seeds to get a lot of DNA!

6. What enzyme is found in meat tenderizer? 13. Why are peas used in this experiment? Are they the best
source of DNA?
The two most common enzymes used in meat tenderizer are
Bromelain and Papain. These two enzymes are extracted from Peas are a good source of DNA because they are a seed. But, we
pineapple and papaya, respectively. They are both proteases, also chose the pea for historical reasons. Gregor Mendel, the father
meaning they break apart proteins. Enzymatic cleaning solutions of genetics, did his first experiments with the pea plant.
for contact lenses also contain proteases to remove protein build-
up. These proteases include Subtilisin A (extracted from a bacteria) 14. How does the experiment compare when using animal cells
and Pancreatin (extracted from the pancreas gland of a hog). instead of plant cells?

7. How much pineapple juice or contact lens solution should I The DNA molecule is structurally the same in all living things,
use to replace the meat tenderizer? including plants and animals. That being said, the product obtained
from this extraction protocol may look slightly different depending
on whether it was extracted from a plant or an animal. For Unfortunately, a microscope will not allow you to see the double
example, you may have more contaminants (proteins, helical structure of the DNA molecule. You'll only see a massive
carbohydrates) causing the DNA to appear less string-like, or the mess of many, many DNA molecules clumped together. In fact, the
amount of DNA that precipitates may vary. width of the DNA double helix is approximately one billionth of a
meter! This is much too small to see, even with the most powerful
15. What sources might I use to extract DNA from animal microscope. Instead, a technique called X-ray crystallography can
cells? be used to produce a picture of the DNA molecule. It was by
looking at such a picture (taken by Rosalind Franklin) that James
Good sources for animal cells include chicken liver, calf thymus, Watson and Francis Crick were able to figure out what the DNA
meats and eggs (from chicken or fish). molecule looks like.

16. Why do peas require meat tenderizer, but wheat germ does
not?

We at the GSLC have done a fair amount of testing with the split
pea protocol and the wheat germ protocol. We have found no
difference in the "product" (nucleic acids) that is observable,
whether using meat tenderizer or not. So, the step was left out of
the wheat germ protocol, but kept in the split pea protocol just for
fun.

Even though it's not necessary, it may be doing something we can't

Real-life Applications of the Science

Yes, in theory. The same basic materials are required, but the
protocol would need to be scaled down (using smaller volumes of
water, soap and alcohol). This is because you're not likely starting
the protocol with the required amount—1/2 cup—of human cells!
That means that you will not extract an amount of DNA large
enough to visualize with the naked eye. If you wanted to see it, you
would need a centrifuge to spin down (to the bottom of the tube)
the small amount of DNA present in the sample.

18. What can be done with my extracted DNA?

This sample could be used for gel electrophoresis, for example, but
all you will see is a smear. The DNA you have extracted is
genomic, meaning that you have the entire collection of DNA from
each cell. Unless you cut the DNA with restriction enzymes, it is
too long and stringy to move through the pores of the gel.

A scientist with a lab purified sample of genomic DNA might also

19. How is DNA extraction useful to scientists? When do they

The extraction of DNA from a cell is often a first step for scientists
who need to obtain and study a gene. The total cell DNA is used as
a pattern to make copies (called clones) of a particular gene. These
copies can then be separated away from the total cell DNA, and
used to study the function of that individual gene.

Once the gene has been studied, genomic DNA taken from a
person might be used to diagnose him or her with a genetic disease.
Alternatively, genomic DNA might be used to mass produce a gene
or protein important for treating a disease. This last application
requires techniques that are referred to as recombinant DNA
technology or genetic engineering.

20. Can I use a microscope to see the DNA that I extract?