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DNA Extraction from Bacteria DNA carries in its molecular structure the genetic information for cell development and behavior. Consequently, all living cells contain DNA. DNA can be isolated from cells of any plant, animal, or microorganism. In this laboratory procedure, you will isolate DNA from Escherichia coli. Supplies required – (Students should work in pairs or groups) • 1 capped test tube containing Escherichia coli • detergent solution (diluted clear shampoo or 10% SDS) • 1 capped test tube of chilled 95% ethanol. You can find this in the freezer of the refrigerator at the back of the lab. For best results, this ethanol should be as cold as possible. • meat tenderizer • 1 flat toothpick • 1 Pasteur pipette or medicine dropper with a rubber bulb attached Step 1. Add about 1 inch of detergent solution to the tube containing the bacteria. Cap the tube; invert slowly several times to gently mix the solution without foaming. Place the tube in a hot water bath at 55-60°C for 10 minutes. Detergent and heating dissolve the fats in the walls of the bacteria causing the cell to burst, which frees the genetic material. Step 2. Remove the test tube from the hot water bath. Add two flat toothpicks full of meat tenderizer to the tube, cap the tube, mix gently but thoroughly, and return the tube to the 55-60°C water bath for 20 minutes. Meat tenderizer contains papain, an enzyme that breaks down any proteins that may be attached to the DNA. Step 3. Remove the tube from the hot water bath. Add cold alcohol to the test tube (about 2/3 full) to create an alcohol layer on top of the bacterial solution. Do this by slowly pouring the alcohol down the inside of the test tube with a Pasteur pipette or medicine dropper. DO NOT MIX! DNA is soluble in water but not alcohol. When alcohol is added to the mixture, all the components of the mixture, except for the DNA, stay in solution while the DNA precipitates out into the alcohol layer. Step 4. Let the solution sit for 2-3 minutes without disturbing it. It is important not to shake the test tube. Watch the white DNA precipitate out into the alcohol layer. DNA has the appearance of white mucus.