DNA Extraction of Peas Procedures: 1. In a medicine cup, measure out 5 ml of the...

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DNA Extraction of Peas Procedures: 1. In a medicine cup, measure out 5 ml of the Pea cell solution. 2. Measure out 1.0 ml of liquid Soap with a pipette and add it to the Pea cell suspension. Discard the pipette. Cap the tube tightly and then rotate or invert it gently so as to avoid excessive bubbling. Repeat this rotation several times over a 5 minute period. You may notice that the suspension becomes more viscous as the membranes are lysed. DNA is found inside the nucleus of the cell and detergent breaks down the lipid cell membrane. 3. Add a pinch of Enzymes (meat tenderizer) to the test tube and stir gently. Be careful, if you stir too hard you'll break up the DNA, making it harder to see. Enzymes break down the protein bonds. DNA is molded, folded, and protected by proteins. The enzymes in meat tenderizer cut the proteins away from the DNA. 4. Loosen the cap on the test tube. Stand the tube containing the Pea lysate in a hot water bath preheated to 60-65°C for 30 minutes. 5. Remove the lysate from the water bath. Allow the lysate to cool to room temperature. In a medicine cup, measure out 3-4 ml of ice cold Ethanol 6. Carefully lower the glass rod into the lysate in the spooling tube. With a pipette, add the ice cold ethanol to the tube. To prevent the aqueous and ethanol layers from mixing, tip the tube at a 45° angle, hold the pipet against the side of the tube, and allow the ethanol to flow very slowly down the side of the tube onto the aqueous layer. Discard the pipet. 7. Continue to hold the tube at an angle of 45° so that the maximum interface surface is exposed. Let the straw rest on the bottom of the tube. DNA normally stays dissolved in water, but when in contact with alcohol it precipitates. Slowly rotate the glass rod in a continuous clockwise direction. The physical force of the stringy DNA clumping together as it precipitates pulls more strands along with it as it rises into the alcohol. Avoid touching the sides of the tube. Fibers of DNA will attach to the rod at the interface of the two layers. Continue twirling the straw for several minutes until a visible mass of DNA has attached to the rod. Alcohol layer Interface between layers Aqueous layer Transcription and Translation 4. What is the purpose of the hand soap in the experiment? 5. What is the purpose of the heat during the experiment? 6. What is the purpose of the alcohol during the experiment? 7. What does the DNA look like after completion of the experiment? DNA Extraction of Peas Procedures: 1. In a medicine cup, measure out 5 ml of the Pea cell solution. 2. Measure out 1.0 ml of liquid Soap with a pipette and add it to the Pea cell suspension. Discard the pipette. Cap the tube tightly and then rotate or invert it gently so as to avoid excessive bubbling. Repeat this rotation several times over a 5 minute period. You may notice that the suspension becomes more viscous as the membranes are lysed. DNA is found inside the nucleus of the cell and detergent breaks down the lipid cell membrane. 3. Add a pinch of Enzymes (meat tenderizer) to the test tube and stir gently. Be careful, if you stir too hard you'll break up the DNA, making it harder to see. Enzymes break down the protein bonds. DNA is molded, folded, and protected by proteins. The enzymes in meat tenderizer cut the proteins away from the DNA. 4. Loosen the cap on the test tube. Stand the tube containing the Pea lysate in a hot water bath preheated to 60-65°C for 30 minutes. 5. Remove the lysate from the water bath. Allow the lysate to cool to room temperature. In a medicine cup, measure out 3-4 ml of ice cold Ethanol 6. Carefully lower the glass rod into the lysate in the spooling tube. With a pipette, add the ice cold ethanol to the tube. To prevent the aqueous and ethanol layers from mixing, tip the tube at a 45° angle, hold the pipet against the side of the tube, and allow the ethanol to flow very slowly down the side of the tube onto the aqueous layer. Discard the pipet. 7. Continue to hold the tube at an angle of 45° so that the maximum interface surface is exposed. Let the straw rest on the bottom of the tube. DNA normally stays dissolved in water, but when in contact with alcohol it precipitates. Slowly rotate the glass rod in a continuous clockwise direction. The physical force of the stringy DNA clumping together as it precipitates pulls more strands along with it as it rises into the alcohol. Avoid touching the sides of the tube. Fibers of DNA will attach to the rod at the interface of the two layers. Continue twirling the straw for several minutes until a visible mass of DNA has attached to the rod. Alcohol layer Interface between layers Aqueous layer Transcription and Translation 4. What is the purpose of the hand soap in the experiment? 5. What is the purpose of the heat during the experiment? 6. What is the purpose of the alcohol during the experiment? 7. What does the DNA look like after completion of the experiment?

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