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Jo Ann Lane
1994 Woodrow Wilson Collection
Overview
To most students of biology, DNA is an abstraction. You can memorize the names and structures
of the nitrogenous bases and know all about the history of DNA's discovery, but until you
actually handle DNA, it remains a strange and mysterious substance.
The purpose of this laboratory is to give you firsthand experience with DNA by isolating it from
plant tissue. You will start with whole onions and end with a relatively pure preparation of DNA,
containing literally millions of genes. Once isolated, the DNA can be stored in alcohol or dried
out. It will actually be possible for you to hold in your hands the key to an organism's
development and structure.
Objectives
1. To become familiar with the physical properties of DNA by isolating it from living tissue
2. To learn the purpose of each step in the isolation procedure as it relates to the physical
and biochemical characteristics of the genetic material
Materials and Equipment
The following materials can be shared by a group of students:
• blender
• 60û C water bath
• thermometer
• ice bucket
• balance (0.1 g scale)
• 95% ethanol kept on ice
The following materials are needed for each student or pair:
• plastic gloves
• 100 ml homogenization medium cheesecloth (4 thicknesses
2-100 ml graduated cylinders (one kept on
• cutting board ice)
• medium onion 250 ml beaker
• knife 500 ml beaker
1000 ml beaker
• weighing boat glass stirring rod
• funnel
2. What did you learn about the properties of DNA during this laboratory period?
3. What structural characteristics of DNA allows it to be spooled out on a glass rod? Why is
it not possible to spool out precipitated proteins? (Hint: Compare the relative lengths of
DNA and protein molecules.)
Teacher Information
1. Preparation of homogenization medium:
sodium laural sulfate (SDS or SLS) 50.000 g
sodium chloride 8.770 g
sodium citrate 4.410 g
ethylenediamine tetraacetic acid (EDTA) 0.292 g
2. Add distilled water to make 1 liter of solution. Do not place in refrigerator or the SDS
will turn the solution an opaque white color. If the homogenization medium gets cold at
any time, it will turn white, but this will not affect its function.
3. Ethanol must be cold for this procedure to work. Place a bottle of ethanol and one
graduated cylinder for each lab group in a freezer overnight. Be sure the cap is loose and
the bottle is not completely full. Place the bottle on paper towels. It will not freeze. Just
before lab dispense the ethanol into smaller containers and put on ice. Or you can fill one
smaller bottle for each lab group and pass out the bottles and graduated cylinders directly
from the freezer or from a cooler filled with ice just before the students will use them.
4. Have the students wear gloves and tell them not to touch the inside of containers because
DNAse enzymes from their hands will break the DNA into small fragments so that it will
not spool at the end of the lab. Rinse all glassware with distilled water.
5. Stress with the students that they must follow the directions carefully since the
temperatures and timings are crucial to the procedure.
6. Scoring the end of the glass rods with sandpaper will help the DNA adhere to the rod
while spooling. Do not touch the end of the rod with your fingers.
7. You may have some small vials and 50% ethanol available so that the students may save
their DNA.
8. To save time you may use a blender to chop up the onions and dispense them in 50 g
portions.
9. There is a lot of "waiting time" in this lab, so a worksheet on DNA can be completed
during this time.
10. Wear a mask when massing the sodium laural sulfate-it is very powdery and gets into the
air.