Practical 6.

1
Extracting DNA from onion bulb cells
Safety
The normal safety precautions associated with the use of chemicals and sharps apply.

Apparatus and materials

glass rod
blender
water bath set at 60 C
two 400 cm3 beakers
boiling tube
test-tube rack
funnel and coarse filter paper (or coffee
filter paper)
Pasteur pipette
knife or scalpel
microscope slide

chopping board
onion bulb (Allium sp.)
universal indicator
600 cm3 beaker containing small amount of ice
(as an ice-water bath)
3 g of sodium chloride and 10 cm3 of washing-up
liquid dissolved in 100 cm3 of distilled water
10 cm3 of 95% ethanol stored in a freezer for at
least 12 hours before use
2 cm3 of 1% pepsin
eye protection

Introduction
In this practical, you will extract some DNA from onion cells.

Procedure
1

Cut an onion bulb into small pieces and transfer them to a beaker. Add approximately 100 cm3 of
the salt and detergent mixture, and stir thoroughly.

Place the beaker in a water bath at 60 C for 15 minutes.

The detergent forms complexes around the membrane phospholipids and proteins, causing them to
precipitate out of solution. The sodium ions from the salt shield the negatively charged phosphate
groups of the DNA molecules, causing them to coalesce. At 60 C, nuclease enzymes, which
would otherwise start to fragment the DNA, are partially denatured.

Cool the mixture in an ice-water bath for 5 minutes, stirring frequently.

This slows down the denaturation of DNA that would occur if a high temperature were
maintained.

Pour the mixture into a blender and blend for 5 seconds.

This permits the release of DNA by further degrading the cell walls and membranes. However, the
DNA will be broken up if blending is carried out for more than 5 seconds.

Filter the mixture into a beaker, ensuring that the foam on the surface of the mixture does not
contaminate the filtrate.

Transfer 6 cm3 of the filtrate to a clean boiling tube.

Add four drops of the 1% pepsin solution provided. Mix thoroughly.

Pepsin hydrolyses the proteins in the mixture to amino acids.

Cambridge University Press 2013

Practical 6.1

Slowly pour 9 cm3 of ice-cold 95% ethanol down the side of the tube so that it forms a layer over
the filtrate and enzyme mixture. Leave the tube for a few minutes without disturbing it.

Using a Pasteur pipette, try to draw up some threads of the fibrous material that forms in the cold
ethanol and transfer it to a slide. These threads contain DNA from the onion cells. Add a drop of
universal indicator to confirm that it is acidic.

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