Isolate DNA from a plant tissue Simplified Revision Notes for Leaving Cert Biology
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Isolate DNA from a plant tissue
Materials needed:
Kiwi
Washing up liquid
Table salt
Protease enzyme
Ice cold ethanol
Method:
Chop the kiwi into small pieces.
Add the chopped kiwi to a beaker containing a salt and washing-up liquid solution, then stir.
Chopping the kiwi breaks down the cell wall, increasing surface area.
The detergent dissolves the cell membranes
The salt helps the DNA to clump together.
Place the beaker in a water bath at 60°C for exactly 15 minutes.
This denatures the enzymes that would breakdown the DNA.
Cool the mixture by placing the beaker in an ice-water bath for 5 minutes.
Stops breakdown of DNA.
Pour the mixture into a blender and blend for no more than 3 seconds.
Further breaks down the cell wall, but blend for no more than 3 seconds to avoid DNA degradation.
Carefully filter the mixture into a second beaker.
Transfer some of the filtrate into a boiling tube.
Add 2-3 drops of protease to the boiling tube.
Slowly trickle ice-cold alcohol down the side of the boiling tube.
This separates DNA from the filtrate.
DNA is insoluble in cold ethanol. The DNA will rise to the ethanol-filtrate boundary.
Observe any changes at the interface of the alcohol and the filtrate.
Using a glass rod, gently draw the DNA out from the alcohol layer.
Record the result.
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