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Lab Activity - Extracting DNA: Materials

This document provides instructions for extracting DNA from various plant and animal materials. Key steps include blending the material with salt water, adding detergent to lyse the cells, and then precipitating the DNA by adding alcohol. The long, stringy white clumps that form at the interface of the water and alcohol layers are DNA molecules that have been precipitated out of solution. Storing the extracted DNA in alcohol helps preserve it.

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John Osborne
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163 views

Lab Activity - Extracting DNA: Materials

This document provides instructions for extracting DNA from various plant and animal materials. Key steps include blending the material with salt water, adding detergent to lyse the cells, and then precipitating the DNA by adding alcohol. The long, stringy white clumps that form at the interface of the water and alcohol layers are DNA molecules that have been precipitated out of solution. Storing the extracted DNA in alcohol helps preserve it.

Uploaded by

John Osborne
Copyright
© © All Rights Reserved
Available Formats
Download as PDF, TXT or read online on Scribd
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Lab Activity – Extracting DNA

Online virtual extraction: http://learn.genetics.utah.edu/content/labs/extraction/

DNA can be extracted from anything. Since DNA is the blueprint for life – all living organisms contain
DNA. Suggestions are plant specimens such as peas (even dried peas), strawberries (frozen strawberries
are fine), broccoli, wheat germ, spinach or animal issue such as liver or thymus.

Materials
✔Blender ✔Salt ✔Cold water ✔Strainer/sieve
✔Funnel ✔Test tube ✔2 beakers ✔ Graduated cylinder
✔Meat tenderizer, containing either bromelain (from pineapples) or papain (from papayas)
✔Clear liquid dishwashing detergent (look for sodium laurel sulfate in the ingredients)
✔Cold alcohol (91-95%) ✔Wooden stick for spooling ✔ Glass stirring rod

Method
Put in a blender:
● 1/2 cup of plant or animal material – eg strawberries, peas,
liver, etc (100ml)
● 1/8 teaspoon table salt (less than 1ml)
● 1 cup cold water (200ml) – less if you are using watery
material such as grapes

Why is cold water important? Using ice-cold water and ice-cold alcohol will increase your yield of DNA.
The cold water protects the DNA by slowing down enzymes that can break it apart. Why would a cell
contain enzymes that destroy DNA? These enzymes, called Dnases, are present in the cell cytoplasm (not
the nucleus) to destroy the DNA of viruses that may enter our cells and make us sick. A cell's own DNA is
usually protected from Dnases by the nuclear membrane, but adding detergent destroys that membrane.
The cold alcohol helps the DNA precipitate (solidify and appear) more quickly.
Salty water helps the DNA precipitate (solidify and appear) when alcohol is added.

● Blend on high for 15 seconds. The blender separates the cells


from each other, so you now have a really thin cell soup.
● Pour your cell soup through a strainer or sieve into another
beaker.
● Add 2 tablespoons liquid detergent (about 30ml) and swirl to
mix.
● Let the mixture sit for 5-10 minutes.
● Pour the mixture into test tubes or other small glass
containers, each about 1/3 full.
● Add a pinch of enzymes (meat tenderizer) to each test tube
and stir gently with the glass rod. Be careful! If you stir too
hard, you'll break up the DNA, making it harder to see.
The two most common enzymes used in meat tenderizer are Bromelain and Papain. These two enzymes
are extracted from pineapple and papaya, respectively. They are both proteases, meaning they break
apart proteins.

● Tilt your test tube and slowly pour alcohol (70-95% isopropyl or ethyl alcohol) into the tube down the
side so that it forms a layer on top of the soupy mixture. Pour until you have about the same amount
of alcohol in the tube as the mixture of cells, detergent and meat tenderizer.

Alcohol is less dense than water, so it floats on top. Look for


clumps of white stringy stuff where the water and alcohol
layers meet.

What is the white stringy stuff? DNA is a long, stringy molecule. The salt that you added in step one
helps it stick together. So what you see are clumps of tangled DNA molecules!

DNA precipitates when in the presence of alcohol, which means it doesn't dissolve in alcohol. This causes
the DNA to clump together when there is a lot of it. And, usually, cells contain a lot of it!

DNA normally stays dissolved in water, but when salty DNA comes in contact with alcohol it becomes
undissolved. This is called precipitation. The physical force of the DNA clumping together as it
precipitates pulls more strands along with it as it rises into the alcohol.

You can use a wooden stick or a straw to spool and collect the DNA. If you want to save your DNA, you
can transfer it to a small container filled with alcohol.

The white stringy stuff is actually a mixture of DNA and RNA. 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 smaller pieces,
making the DNA harder to see. If it disappears it's likely because enzymes are still present that are
breaking apart the DNA in your sample.

Cells with more chromosomes contain relatively more DNA, but the difference will not likely be
noticeable. The amount of DNA you will see depends more on the ratio of DNA to cell volume.
(Strawberries are octoploid - so there is lots of DNA to see)

Videos:
How to extract DNA from a Strawberry -
http://www.youtube.com/watch?v=DGnmofoKmHQ&feature=related

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