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DNA Extraction

DNA Extraction. Biology Agriculture. DNA Source. Green Peas. Blender. ½ cup of DNA (peas) A large pinch of table salt (less than 1/8 teaspoon) Twice as much cold water as the DNA source (about 1 cup) Blend on high for 15 seconds. Strainer.

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DNA Extraction

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  1. DNA Extraction Biology Agriculture

  2. DNA Source • Green Peas

  3. Blender • ½ cup of DNA (peas) • A large pinch of table salt (less than 1/8 teaspoon) • Twice as much cold water as the DNA source (about 1 cup) • Blend on high for 15 seconds.

  4. Strainer • Pour your thin pea-cell soup through a strainer into another container.

  5. Detergent • Add about 2 (tablespoons) of detergent, swirl to mix. • Let the mixture sit for 5-10 minutes.

  6. Why add detergent? • Blending separated the pea cells. • But each cell is surrounded by a sack (the cell membrane). DNA is found inside a second sack (the nucleus) within each cell. • To see the DNA, we have to break open these two sacks.

  7. Why add detergent? • We do this with detergent. • Think about why you use soap to wash dishes or your hands. To remove grease and dirt, right?

  8. Why add detergent? • Soap molecules and grease molecules are made of two parts: • Heads, which like water • Tails, which hate water.

  9. Why add detergent? • Both soap and grease molecules organize themselves in bubbles (spheres) with their heads outside to face the water and their tails inside to hide from the water.

  10. Why add detergent? • When soap comes close to grease, their similar structures cause them to combine, forming a greasy soapy ball.

  11. Why add detergent? • A cell's membranes have two layers of lipid (fat) molecules with proteins going through them.

  12. Why add detergent? • When detergent comes close to the cell, it captures the lipids and proteins.

  13. Meat Tenderizer • Pour the mixture into test tubes or other small glass containers, each about 1/3 full. • Add a pinch of enzymes to each test tube and stir gently. Be careful! If you stir too hard, you'll break up the DNA, making it harder to see.

  14. What is an enzyme? • Enzymes are proteins that help chemical reactions happen more quickly. Without enzymes, our bodies would grind to a halt.

  15. What is an enzyme? • In this experiment, the enzyme we are using comes from meat tenderizer and cuts proteins just like a pair of scissors.

  16. What is an enzyme?

  17. What is an enzyme? • After the detergent step, the last question was: what do you have now in your pea soup? • The cell and nuclear membranes have been broken apart,as well as all of the organelle membranes.

  18. What is an enzyme? • So what is left? • Proteins • Carbohydrates (sugars) • DNA

  19. What is an enzyme? • The DNA in the nucleus of the cell is molded, folded, and protected by proteins. The meat tenderizer cuts the proteins away from the DNA.

  20. Mixing Together • Tilt your test tube and slowly pour rubbing alcohol) into the tube • Pour it down the side so that it forms a layer on top of the pea mixture. • Pour until you have about the same amount of alcohol in the tube as pea mixture.

  21. Extracting DNA • DNA will rise into the alcohol layer from the pea layer. • Use a wooden stick draw the DNA into the alcohol.

  22. What is the stringy stuff? • Alcohol is less dense than water, so it floats on top. • Since two separate layers are formed, all of the grease and the protein that we broke up in the first two steps and the DNA have to decide which layer to go to?"

  23. What is the stringy stuff? • In this case, the protein and grease parts find the bottom, watery layer the most comfortable place, while the DNA prefers the top, alcohol layer. • DNA is a long, stringy molecule that likes to clump together.

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