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DNA Exraction Lab
DNA Exraction Lab
A would look like small thin strands. DNA is found in the nucleus and mitochondria. If avocado is put through the steps of DNA extraction, then the DNA will precipitate out of the ethanol solution. Independent Variables: Type of living organism Dependent Variable: DNA extraction Materials:
Heavy duty Ziploc bag 1 strawberry 10mL DNA extraction buffer (soapy, salty water) Cheesecloth Funnel 50mL vial / test tube Glass rod, inoculating log, or popsicle stick 20 mL ethanol
Procedure: 1. Place one strawberry in a Ziploc bag. 2. Smash/grind up the strawberry using your fist and fingers for 2 minutes. Careful not to break the bag!! 3. Add the provided 10mL of extraction buffer (salt and soap solution) to the bag. 4. Kneed/mush the strawberry in the bag again for 1 minute. 5. Assemble your filtration apparatus as shown to the right. 6. Pour the strawberry slurry into the filtration apparatus and let it drip directly into your test tube. 7. Slowly pour cold ethanol into the tube. OBSERVE _ 8. Dip the loop or glass rod into the tube where the strawberry extract and ethanol layers come into contact with each other. OBSERVE _ (TAKE A PICTURE) 9. Repeat steps 1-8 with something else that you think will have DNA in it to see if you can get the DNA to precipitate out of that. BEFORE YOU DO THIS PART, MAKE A
HYPOTHESIS. 10. After the class has finished, take note of which items were able to have their DNA extracted. 11. Put all of the results from second round into a data table. Data: Living Organism Strawberry Kiwi Pepper Canned Carrots Banana Spinach Watermelon Avocado Apple Cucumber Tomato Rating of how visible the DNA was 5 1 3.5 5 3.5 5 1 3 4 1 4
Graduated Cylinder
Test tube
Funn el
Analysis: My Hypothesis was correct because after the ethanol and avocado settled the DNA was visible. At first the DNA was not visible and you can see this in the picture above, but after it settled the DNA
became visible. I noticed that the fattier living organisms and the living organisms with higher water content had less obvious DNA. During the lab at first I thought that the DNA was not extracted from the avocado but this issue was fixed when the mixture settled. One variable that I did not account for in the lab that would have made the DNA extraction more obvious was the fat content to soap ratio. If I were to do this lab again, I would set a ratio of the fat content to soap and salt to strength of cell wall that I would keep constant between every living organism. The purpose of the initial smashing and grinding of the strawberry was to break open the cells, the purpose of mushing the strawberry was to break up the proteins and dissolve the cell membranes, the purpose of filtering the strawberry slurry through the cheesecloth was to separate the components of the cells, and the purpose of the addition of ethanol to filter the extract was to precipitate the DNA from the solution. The independent variable in this lab is the type of living organism used and the dependent variable in this lab is whether the DNA was able to be extracted because the extraction of DNA depended on the type of living organism used in the lab. The DNA looked like long strands of very fine thread until it stuck to itself and formed a blob that felt like a wet piece of crumpled up paper. My knowledge of how one cells DNA is about three meters long and a thin double-helix can be related to this observation because the strand were long although not quite three meters, the fact that they got clumped together so easily could be related to the double-helix because double-helixes can bend and fold easily, and the thinness of the DNA could be related to how thin the strands I saw were. In the final step when we added ethanol to the strawberry extract, the DNA was visible and we were able to take it out of the test tube because the DNA wasnt soluble in the ethanol so it precipitated from the ethanol solution rather than dissolving. I was correct on my hypothesis on the second trial because the DNA from the avocado did end up precipitating out of the ethanol. All of the objects that the class precipitated DNA from the ethanol but some of the DNA precipitated more obviously because some objects had higher water content or a stronger cell wall or a higher fat content. Conclusion: The purpose of the lab was to extract and view DNA and the data collected supports this purpose because the ratings are based off of how well the DNA was extracted and how visible the DNA was. The data from the lab can be related to the real world because it shows that living organisms with a lot of fat or a very strong cell wall are harder to get into. The data could have been more conclusive if we had tried animal cells rather than just plant cells and it is unknown whether the soapy, salty water and the ethanol would have been the same materials to extract DNA from an animal cell in addition to plant
cells. The statement, A person cannot see a single cotton thread 100 feet away, but if you wound thousands of threads together into a rope, it would be visible much farther away, is analogous to our DNA extraction because a single strand of DNA is microscopic but when we had DNA from many cells from one living organisms, the DNA was visible. One reason that it is important for scientists to be able to extract and view DNA, is to make advancements in gene therapy. Another reason that it is important for scientists to extract and view DNA is to allow scientists to figure out which conditions and disorders are genetic and hereditary. There is DNA in all of my food, I know because all of my food has some type of living organism in the ingredients whether it is meat wheat, sugar, vegetables, fruits, or something else.