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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.
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.