Exercise 4

1
Exercise 4

• DNA Isolation, Restriction Enzyme, Digestion, and
  Gel Electrophoresis

2
Exercise 4 Page 27

1. Write your group on the test tube
2. With graduated cylinder, get 10-15ml of
   detergent/buffer solution from the refrigerator
   and add it to your test tube
3. Stir vigorously with the large glass stirring
   rod. Be careful not to break the rod or test
   tube
4. Answer the first question on page 28
5. Place the test tube in the warm water bath for 20
   minutes.

3
DNA

• What does DNA stand for?
• Deoxyribonucleic acid
• Where do you find DNA?
• In the nucleus
• What is DNA?
• Genetic information
• What do we use DNA testing for?
• Paternity, crimes, identification, etc
• Where can you get DNA samples?
• Blood, hair, saliva, skin, etc

4
DNA

• What is DNA made of?
• Nucleic acid (the 4th carbon compound)
• Two strands of nucleotides
• Sugars, phosphates, nitrogenous bases
• What are the nitrogenous bases?
• Adenine
• Cytosine
• Guanine
• Thymine
• Which nitrogenous bases associate?
• Adenine Thymine
• Cystosine Guanine

5
DNA

• Restriction enzymes enzymes that break down DNA
  at certain sequence
• Eco RI cuts at GAATTC

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Exercise 4 Page 28

1. Gently stir test tube with large glass rod
2. Place a funnel into the 50ml beaker
3. Place a piece of cheesecloth over funnel
4. Pour your test tube solution into the funnel and
   allow to filter into beaker (need 10-15ml of
   solution)
5. Remove funnel and cheesecloth
6. With graduated cylinder, get 5ml of meat
   tenderizing solution from the refrigerator and
   add to the beaker
7. Use the small stirring rod to mix GENTLY
8. Allow mixture to sit on your desk for 5 minutes
9. Answer questions in the book while you wait

7
Exercise 4 Page 28-29

1. With graduated cylinder, get 10-15ml of 95
   ethanol.
2. Slightly tilt your beaker while SLOWLY adding
   ethanol
3. Wait 2 minutes
4. Place the small glass rod straight down into the
   beaker until it touches the bottom
5. Gently twist the rod in one direction. A white
   thread-like precipitate (DNA) will wrap around
   the glass rod as you stir
6. Continue turning until you have spooled the DNA
   onto rod
7. Carefully tilt the rod and lift out the DNA
8. Hold the glass rod in the air for 1 minute to let
   the DNA dry
9. While waiting, another team member needs to get a
   tiny test tube and add about 1/3 full or 1 of
   water.
10. Add DNA to small test tube

8
Exercise 4 Page 30

1. Send ONE team member up front to learn how to
   pipette. (Clean up while your classmates are
   learning how to pipette. DO NOT throw away your
   DNA)
2. You have a small blue tube and a small yellow
   tube. Label the top of the lid. Blue is A and
   yellow is B
3. On the side of each tube write your group G1
   and your section. Mon D, Tues H, Wed L,
   Thurs Q
4. Pipette 18µl of DNA solution into blue tube and
   18µl into yellow tube.
5. Use pipette tip to mix
6. Bring tube A (blue) to me for restriction enzyme
   buffer and enzyme
7. Place tubes into the incubator

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Photsynthesis

• Exercise 8 - Record appearance and root/stem
  measurements and replace solutions if needed.
  Page 64
• Exercise 7 Record height of each plant on Page
  58 for both wild type and rosette. Add one drop
  of water to each leaf of your A plants and add
  one drop of gibberellic acid to each leaf of your
  B plants