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DNA Extraction Lab

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Introduction

Biology Lab: Extraction of Crude DNA from Plant and Animal Tissues By: X Class: IB1 Date: Friday, May 15, 2009 Background and Purpose: - In this experiment you will be extracting DNA from two sources: animal tissue (liver tissue) and plant tissues (kiwi). Two procedures will be followed and instructions must be followed precisely in order to obtain appropriate results. - The purpose of this experiment is to strengthen and acquire skills of measuring, observing, predicting, collecting and recording data. We will also learn how different solutions and substances lead to the extraction of DNA. Procedure 1: Extracting DNA from Fresh Sheep Liver Materials: - Fresh Sheep Liver - Liquid Soap - Scalpel or Dissecting Scissors - 95% ethanol - Blender - 0.9% NaCl solution (saline) - Graduated Cylinders - Glass Stirring Rod - Droppers - Beakers - Cheesecloth Procedure and Explanations: 1- Cut a piece of liver about 2 cm long and place it with 30 ml of saline solution in a Blender. ...read more.

Middle

5- Slowly stir the mixture with the glass stirring rod. A Whitish substance (DNA) will form where the alcohol and liver mixture meet. 6- Watch as DNA precipitates up through the ethanol. Twirl the glass rod slowly in the mixture. DNA strands will accumulate and clump together on the rod, forming a visible mass. Record you results. Observations: - Light brownish mixture of liver-ethanol - Small and thick whitish accumulations (DNA) - Small bubbles forming around the DNA Procedure 2: Extracting Crude DNA from Kiwi Fruit Materials: - 1 Kiwi Fruit - Ice cold Ethanol - Water Bath set at 60 oC - Glass Rod - Crushed Ice - Detergent (washing up liquid) - Scalpel - 3% NaCl solution (saline) - Filter Paper - Funnels - Beakers/Flasks - Test Tubes and Rack Procedure and Explanations: 1- Chop Kiwi into small cubes, side about 0.5 cm. ...read more.

Conclusion

DNA is now found in the solution that has been filtered. 6- Transfer DNA solution into a test tube. Pour ice-cold ethanol carefully down the side of the tube on the surface to form an ethanol layer above the extract. The reason of why we use ice-cold ethanol is for precipitating or solidifying the DNA, since DNA will most likely precipitate if it is cooler. The cold temperature of the ethanol just increases the amount of DNA precipitated. 7- Agitate the Liquid by twisting a fine glass rod gently where the two liquids meet. This will gently cause the bubbles and DNA to precipitate to the top of the test tube. 8- Small Bubbles will attach to the DNA strands as they move up through ethanol. They are seen as opaque Stands. Observations: - Bubbles attached to the DNA strands as the moved up the ethanol. - The DNA strands were smaller than the animal tissue DNA. - Solution in test tube was clear. - Small accumulation of DNA on surface. Sources: http://biotech.biology.arizona.edu/labs/DNA_Kiwifruit_teacher.html http://www.biologycorner.com/worksheets/DNA_extraction.html http://biology.arizona.edu/sciconn/lessons2/Vuturo/vuturo/dna.htm http://www.exploratorium.edu/ti/human_body/dna.html http://www.funsci.com/fun3_en/dna/dna.htm ...read more.

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