Lab4 Lab Report

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University of Texas, Rio Grande Valley *

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Biology

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Dec 6, 2023

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Lab #4 DNA Extraction from Cells Date: 10/12/2023 OBJECTIVES To describe where DNA is located in a cell To explain the procedures required to release DNA from a cell To observe the extraction of genomic DNA from a cell INTRODUCTION In Lab 4, DNA Extraction from Cells, we will be extracting the DNA of a banana. We will be breaking its nuclear membrane and viewing its chromosomes. MATERIALS 20 mL DNA extraction buffer (15g NaCl/1L distilled water) 1 mL of liquid dishwashing detergent Ice-cold 95% ethanol or 95%isopropyl alcohol 1/3 banana Ziplock plastic bag Large Test tube Cheese cloth Glass rods 200 mL beaker Elastic PROCEDURE 1. Put 1/3 of a banana into the ziplock bag, squeezing out all air. 2. Gently mash the banana to a pulp for 2 minutes. 3. Open the bag and add 20 mL of extraction buffer into ziplock bag. 4. Filter the mixture through a cheese cloth, save the filtrate (liquid coming through). 5. Pour 10 mL of the filtrate into a test tube. 6. Add 1 mL of detergent (this breaks apart the cell membrane and nuclear membrane) and gently mix solution with a glass stirring rod without creating foam. 7. Immediately add 20 mL of ICE COLD ethanol to test tube by slightly tilting the test tube and SLOWLY pouring it down the side so that a layer of ethanol rests on top of the filtrate/detergent solution (DNA is not soluble in ice cold ethanol so when ethanol is added to the mixture all components of the mixture except DNA stay in solution, while DNA precipitates out). 8. Let the ethanol sit 2-3 minutes without disturbing it (bubbles should form and DNA should precipitate out of the solution at the interface). 9. Gently swirl the DNA using a wooden splint and the solution should look like whitish mucous. 10. The DNA may be lifted out carefully and dried on an absorbent towel. POST-LAB QUESTIONS 1. What was the purpose of mashing up the banana ? It breaks down the tough walls of the plant cells and breaks tissues.
2. What does the extraction buffer do ? It breaks the line between proteins. 3. What does the cheese cloth do ? It removes the solid parts. 4. What happened when you added the detergent to the filtrate ? Detergent dissolves the lipids in the cell and nuclear membrane. 5. What happened when you added the alcohol to the filtrate ? When alcohol is added to the mixture all components of the mixture except DNA stay in solution, while DNA precipitates out. 6. What did the DNA look like ? It kind of looks like whitish mucous. 7. A person cannot see a single cotton thread 15 feet away but if you wound together thousands of cotton threads into a rope would it be visible at the same distance? How is this statement an analogy to our DNA extraction ? Seeing one single DNA is difficult just like trying to see a single strand of cotton thread. Which is why when the DNA is all clumped up together it’s easier to see it. 8. Is DNA found in all living organisms or just once living organisms? Is DNA found in all cells or just some cells? What are some examples of cells that don’t contain DNA in the human body ? DNA is found in all living organisms. However, DNA is not found in all cells for example in skin, hair, and nails. 9. What structures make it possible for DNA to be seen under a microscope ? Chromosomes
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