DNA Extraction

Pre-lab questions:
1. What do you think the DNA will look like?
Mutilated somehow
2. Where is DNA found?
In the nucleus

Introduction
DNA is the fundamental building block for life. It resides in the nucleus of cells and consists of thousands of genes
that contain the instructions for building the different parts of the cell. DNA is what makes us who we are. It
determines what physical characteristics we have, such as hair and eye color, as well as some of the diseases
we may develop. DNA is a very long molecule, so in order to fit inside the cell, the DNA is highly coiled and
twisted into chromosomes.

A cell is a complex living unit that contains many biological molecules and organelles, each performing a specific
function within the cell. In order to analyze the DNA found at a crime scene, the DNA needs to first be extracted
from the cells. This means that it must be separated from all of the other chemicals and materials inside the cell.
Equipment

●​ Computer with Internet access ●​ Small paper or plastic cups (e.g., bathroom
●​ 95% Alcohol (kept ice cold) cups)
●​ DNA extraction buffer ●​ 15 mL Screw-cap conical bottom plastic
●​ (2) 8cm Squares of cheesecloth centrifuge tubes
●​ Distilled water ●​ Plastic transfer pipettes
●​ Cups of crushed ice ●​ wooden popsicle stick
●​ Strawberries
●​ Ziploc freezer bags (small, quart size)

Procedure
In this lab you will isolate DNA from plant and animal cells. In order to extract DNA from cells, scientists take
advantage of the unique chemical properties of the DNA. Plant cell walls are made of cellulose, which is a tough,
insoluble material that makes plants sturdy. It can be broken through physical actions. In this lab you will mash the
strawberries in order to break open the cell walls. Cell membranes and nuclear membranes are made up of fats.
Detergents wash away these fats, just like they would grease in a pan. In this lab you will treat the mashed
strawberry cells with detergent to dissolve the cell membranes and nuclear membranes to release the DNA. Once
the cells are broken open, the DNA and cell debris are all mixed together. Because the DNA is in solution, you can
separate the components by filtering the mixture through cheesecloth. Finally, you can separate the DNA from
solution with alcohol, since DNA does not dissolve in alcohol. It is very important that you follow the procedure
carefully. Otherwise, your DNA sample will be contaminated with proteins and other cell components. Remember to
follow all the safety instructions provided by your teacher.

DNA Isolation from Fruit:

1.​ Put on the appropriate personal protective equipment, including safety gloves and goggles. Tie back any
loose hair.
2.​ Get ½ a strawberry from your instructor. Place in your bag and make sure all of the air is out of the bag and
mash the strawberry very well. Do not smash the strawberry or the bag will break.
3.​ Bring your bag to your teacher and get 15 mL of DNA extraction buffer. Seal the bag and mush together for 1
minute.
 4.​ Place the square of cheesecloth over a small paper or plastic cup. Make sure to create a pocket deep
enough to hold the mixture and cover the rim of the cup. Secure with a rubber band if needed.
5.​ Pour the strawberry mixture slowly into the filter. Make sure NOT to overfill. Pour in only as much as needed
to fill the filter at one time.
6.​ Once the mixture has filtered through, carefully pour 2 mL of the filtered contents into a clean 15 mL conical
tube.
7.​ Hold the 15 mL conical tube at an angle. Using a transfer pipet, carefully and slowly add 5 mL of cold 95%
ethanol down the side of the tube. The mixture will sink to the bottom of the tube and alcohol will sit on top.
Do NOT mix. Allow the tube to sit undisturbed for two or three minutes.
8.​ Watch closely as the DNA will appear as a clear-to-whitish, stringy clump where the ethanol layer meets the
strawberry extract layer. Tiny bubbles in the ethanol layer will appear where the DNA precipitates.
9.​ Dip the wooden popsicle stick into the tube and twirl it around gently to collect some DNA. Stir the DNA gently
at the interface between the ethanol and strawberry extract layer to extract more fibers.
10.​Carefully remove the popsicle stick from the tube and observe the DNA. Record all observations in your
laboratory journal. Take a picture.
11.​Wash your bag so it can be reused. Put the rubber band back into the big cup. The cheesecloth, small dixie
cup, and popsicle stick can be thrown away.
DNA Isolation from Saliva - We will only be completing Plant Cell DNA Extraction
1.​ Scientists can use a procedure similar to the one found below to extract DNA from the cells of victims or
potential suspects, either in saliva or other bodily fluids. They must be very precise as they work to prevent
contamination of the final sample.
2.​ Get a salt solution from your teacher and rinse your mouth for one minute with the salt solution. Make sure to
use your teeth to scrape the inside of your cheeks. This will release the cells into the solution. Spit the salt
solution containing your cheek cells back into your cup.
3.​ Pour the salt solution with saliva/cheek cells into a cup (a new cup, do not use your strawberry cup).
4.​ Bring your cup to your teacher and add the detergent solution. Gently rock the mixture back and forth for 2
minutes. Add 8mL of this solution to a clean conical tube.
5.​ Hold the 15 mL conical tube at an angle. Using a transfer pipet, carefully and slowly add 3 mL of cold 95%
ethanol down the side of the tube. The mixture will sink to the bottom of the tube and alcohol will sit on top.
Do NOT mix. Allow the tube to sit undisturbed for two or three minutes.
6.​ Watch closely as the DNA will appear as a clear-to-whitish, stringy clump where the ethanol layer meets the
cheek cell extract layer. Tiny bubbles in the ethanol layer will appear where the DNA precipitates.
7.​ Dip the glass stirring rod or wooden popsicle stick into the tube and twirl it around gently to collect some DNA.
Stir the DNA gently at the interface between the ethanol and saliva layer to extract more.
8.​ Carefully remove the stirring rod or popsicle stick from the tube and observe the DNA. Record all
observations in your laboratory journal.
9.​ Clean up your lab station and wash your hands. Wash your tube and reset your station to match what you
walked into.
10.​Answer the Conclusion questions.

DNA from Strawberry Observations DNA from Saliva Observations

Slimy stringy texture wrapping the stick.

Strawberry DNA Picture Saliva DNA Picture

Conclusions and Analysis

2. It is important that you understand the steps in the extraction procedure and why each step was necessary. Each
step in the procedure aided in isolating the DNA from other cellular materials. Match the procedure with its function:

PROCEDURE ​ ​ ​ ​ ​ ​ FUNCTION
A. Filter strawberry slurry through cheesecloth​ __C_ To precipitate DNA from solution
B. Mush strawberry with salty/soapy solution​ _B__ Separate components of the cell
C. Initial smashing and grinding of strawberry​ __A_ Break open the cells
D. Addition of ethanol to filtered extract​ ​ __D Break up proteins and dissolve cell membranes

3. What did the DNA look like? Relate what you know about the chemical structure of DNA to what you observed
today.
The DNA looked stringy and clear.

5. Why is it important for scientists to be able to remove DNA from an organism? List two
Reasons.
Genetic analysis and research
Biotechnology and genetic engineering

6. Is there DNA in your food? __Yes______ How do you know? If it passes the texture test

7. Describe one of the main challenges in extracting DNA from cells found at a crime scene
Degredation or contamination.