The Molecule of Life DNA Indiana University 生命的分子印第安那大学.pptVIP

The Molecule of Life: DNA The Molecule of Life: DNA The purpose of this laboratory exercise is to extract and visualize DNA from fruit. The objectives of the laboratory exercise are: To understand where DNA is found To isolate DNA To understand how DNA is extracted To learn about positive and negative controls What does DNA look like? DNA contains one of four nucleotides: adenine (A), thymine (T), cytosine (C), and guanine (G). A+T or C+G The role of the nucleotides The different nucleotides spell out a code: instructions for the cell Each set of instructions is a gene. A gene is a long series of the four letters (nucleotides) that gives instruction to the cell. Lab Protocol Step 1 – Prepare Materials Solutions Solutions DNA Buffer: Combine 120 mL of dH2O (distilled water) with 1.5 g salt (noniodized), 5 g baking soda and 5 mL dishwashing liquid. Materials Ziploc bag Pipet Bulb (3) 10mL Pipettes Distilled Water Fruit Sample Box of Kimwipes Buffer Cheesecloth Ethanol (95 – 100 percent) Test Tube Rack 15 mL conical tubes Glass rod or wooden stick Metal Spatula Black paper 50 mL conical tube Gloves Scissors Step 2 – Prepare tubes Label the 15 mL conical tubes with your initials or group name. Put on your gloves! Step 3 – Prepare Experimental Samples Weigh out 7.5 g of the fruit from which you will be isolating DNA In the ziploc bag, combine the fruit with 7 mL of dH2O 3 mL of buffer solution Grind the mixture into a fine paste. Why do we crush the fruit? So we can break apart and open the cells. Filter the mixture through at least two layers of cheesecloth into a 50 ml plastic tube. Why add ethanol? Ethanol is less dense than water so it floats on top. All of the proteins we broke up in Step 4 will sink to the bottom; the DNA will float on top. If there is DNA present in any of the samples it should precipitate out in gray clumps that may look like white fine lint fibers. Use a glass rod to spool out the