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http://learn.genetics.utah.edu/content/l abs/extraction/
DNA Source
Green Peas
Blender
cup of DNA (peas) Large pinch of table salt (less than 1/8 teaspoon) Twice as much cold water as DNA source (about 1 cup) Blend on high for 15 seconds
The blender separates the pea cells from each other, so you now have a really thin pea-cell soup.
Strainer
Pour your thin peacell soup through a strainer into another container.
Detergent
Add about 2 tablespoons of detergent, swirl to mix. Let the mixture sit for 5-10 minutes.
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.
What is an enzyme?
Enzymes are proteins that help chemical reactions happen more quickly. Without enzymes, our bodies would grind to a halt.
What is an enzyme?
In this experiment, the enzyme we use comes from meat tenderizer and cuts proteins just like a pair of scissors. You can also use pineapple juice or contact lens cleaning solution as an enzyme.
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.
What is an enzyme?
So what is left?
Proteins Carbohydrates (sugars) DNA
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.
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.
Extracting DNA
DNA will rise into the alcohol layer from the pea layer Use a wooden stick draw DNA into the alcohol
Resources:
University of Utah
Genetic Science Learning Center
HOW TO EXTRACT DNA FROM ANYTHING LIVING
http://learn.genetics.utah.edu/content/labs/ext raction/howto/
Resources:
The rest of these slides are for teacher information, and do not necessarily need to be shown to the class. They are informational text that can be used for deeper understanding of DNA extraction.
Trouble-shooting
1. I dont think Im seeing DNA. What should I be looking for?
Look closely. Your DNA may be lingering between the two layers of alcohol and pea soup. Try to help the DNA rise to the top, alcohol layer. Dip a wooden stick into the pea soup and slowly pull upward into the alcohol layer. Also, look very closely at the alcohol layer for tiny bubbles. Even if your yield of DNA is low, clumps of DNA may be loosely attached to the bubbles.
Trouble-shooting
2. What can I do to increase my yield of DNA?
Allow more time for each step to complete. Make sure to let the detergent sit for at least five minutes. If the cell and nuclear membranes are still intact, the DNA will be stuck in the bottom layer. Or, try letting the test tube of pea mixture and alcohol sit for 30-60 minutes. You may see more DNA precipitate into the alcohol layer over time. Keep it cold. Using ice-cold water and ice-cold alcohol will increase your yield of DNA. The cold water protects the DNA by slowing down enzymes that can break it apart. The cold alcohol helps the DNA precipitate (solidify and appear) more quickly. Make sure that you started with enough DNA. Many food sources of DNA, such as grapes, also contain a lot of water. If the blended cell soup is too watery, there won't be enough DNA to see. To fix this, go back to the first step and add less water. The cell soup should be opaque, meaning that you can't see through it.