DNA Extraction Lab Handout

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Oct 30, 2023

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DNA Extraction Lab 25 Points Cell Theory Matthias Schleiden, Theodor Schwann, and Rudolf Virchow proposed the unified cell theory in the late 1830s. 1. All living things are composed of one or more cells 2. Cell is the basic unit of life 3. All new cells arise from existing cells All cells share four common components: 1. Plasma membrane - Outer covering that separates the cell’s interior from its surrounding environment. 2. Cytoplasm – a jelly-like region within the cell in which other cellular components are found. 3. DNA – the genetic material of the cell 4. Ribosomes – particles that synthesize proteins. Prokaryotic versus Eukaryotic Cells DNA Packaging in a Prokaryotic Cell (image below) DNA is packaged in a very specific way to fit inside the nucleoid region of a prokaryotic cell. Most prokaryotes have a single, circular chromosome in the nucleoid region of the cell. The DNA of E.Coli bacteria is 4.6 million base pairs, which is 1.6 mm if stretched out.
It fits inside a tiny bacterial cell due to the DNA being twisted beyond the double helix, known as supercoiling. DNA Packaging in a Eukaryotic Cell (image on next page) If you stretch a human DNA molecule, end-to-end, it is 2 meters long. In eukaryotes, DNA is present inside the nucleus of the cell. The simplest structure of DNA is the linear, double-helix form.
The linear DNA is wrapped around histone proteins to form nucleosomes. The nucleosomes stack onto each other to form a 30nm-wide chromatin fiber. The fiber is then coiled into a thicker and more compact structure called a chromosome. Plant cells have cell walls, animal cells do not. Experimental Protocol Each Group will be given four test tubes (one test tube per person, two tubes each for what you’re comparing)
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1. Pick two of the DNA sources (blended peas, strawberries, or spinach). 2. Fill approximately 1/3 rd of your test tube with the blended DNA source. 3. Add two drops of diluted dishwashing liquid of any kind to the test tube. Swirl the test tube gently to mix. 4. Let the mixture sit for 10 minutes. 5. Add a pinch of a meat tenderizer or a drop of pineapple juice to the test tube. Swirl the test tube gently to mix. 6. Tilt your test tube and slowly pour 70% ethanol down the side of the test tube, until you have approximately the same volume of alcohol as the blended pea/strawberry or spinach. 7. Alcohol is less dense than water and will float on the top. 8. Look for white clumps of white stringy stuff where the water and alcohol layers meet. THIS IS YOUR DNA! Activity Overview: During today’s lab, you will attempt to extract DNA from a eukaryotic cell using different DNA sources. You will prepare 4 different tubes and visually compare the DNA in the tubes and note down your results. Methods – Experimental Design Each group will develop their own hypothesis and experiment. Your Scientific Question: Your driving question should involve visually observing the amount of DNA in the test tube (rate from 1-4, going from the least amount of DNA to the most DNA in the tube) and the quality of the DNA (is it all in a neat little clump, or is it broken up and floating in the test tube?) Testable questions for the lab include the following examples. i. Do different sources of DNA yield different quantities of DNA?
Write your group’s scientific question(s) here (4 points): What is your hypothesis? (4 points) – cite one source that supports your hypothesis (Use the formula: If EXPLANATION, and METHOD, then PREDICTION) Your Experimental Design : Your experiment must include two different experimental groups What are your proposed experiments? (5 points) (list your two experimental groups; what are you going to do differently in your two groups? The parameters to measure are the same for all tubes – Quantity and Quality of DNA (look at each of the two tubes per question and compare to see which one has more quantity of DNA (more or less) and better DNA quality (DNA is in one big clump or do you see fragments of broken up DNA). Results: Variables tested Test tube DNA Quantity DNA Quality 1 2 1 2
Post-Experiment Questions : 1. Do your results support or reject your hypothesis? Explain. (4 points) 2. What claims can you make based on your results? (2 points) 3. What were your control or experimental variables? (2 points) 4. What methodological improvements could be made to this experiment? Would you do anything differently if you did this experiment again? (4 points)
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