Materials
Fresh thymus
Blender
Beaker
Sugar
Pipet
Centrifuge tube with cap
Bufferin (325mg)
Knife
Graduated cylinders (10ml,100ml)
Epsom salts
Distilled water
Centrifuge
95% ice cold ethanol
15 ml test tubetest tube rack or beaker
Palmolive detergent
Non-iodized salt
Solutions
Prep buffer solution:
57 g granulated sugar
1 buffered aspirin
3 g epsom salts
Add distilled water for a total of 500 ml
10% detergent solution:
90 ml distilled water
10 ml Palmolive detergent
Salt solution:
29.2 g non-iodized salt
Add distilled water for a total volume of 250 ml
Protocol
Cut out a chunk of liver or thymus 1 inch square and place in the blender.
Add 100 -150 ml prep buffer and 10 ml detergent solution to the blender.
Blend for 1 minute or until the mixture is smooth.
Pour the mixture into a beaker.
Transfer 1 ml of the mixture to a centrifuge tube.
Add 2 ml of salt solution, cap, and shake for 2 minutes.
Centrifuge for 7 minutes in a balanced centrifuge.
Carefully remove the tube from the centrifuge and note the two layers:
Lower layer - pellet
*upper layer - liquid (supernatant) and what has the DNA in it.
Pipet or carefully pour the liquid into a clean test tube.
Pour 5 ml ice cold ethanol carefully down the side of the tube to form a layer.
Let the mixture sit undisturbed for a minute or two.
The DNA will float in the alcohol. The DNA of the thymus will be long threads that easily spool.
Modified from: "Generic, All Purpose DNA Extraction from Meat Protocol" Judy Brown
"Mammalian DNA Extraction" Theresa Knapp
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