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THYMUS DNA EXTRACTIONS

Lana Hays

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

  1. Cut out a chunk of liver or thymus 1 inch square and place in the blender.
  2. Add 100 -150 ml prep buffer and 10 ml detergent solution to the blender.
  3. Blend for 1 minute or until the mixture is smooth.
  4. Pour the mixture into a beaker.
  5. Transfer 1 ml of the mixture to a centrifuge tube.
  6. Add 2 ml of salt solution, cap, and shake for 2 minutes.
  7. Centrifuge for 7 minutes in a balanced centrifuge.
  8. 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.
  9. Pipet or carefully pour the liquid into a clean test tube.
  10. Pour 5 ml ice cold ethanol carefully down the side of the tube to form a layer.
  11. Let the mixture sit undisturbed for a minute or two.
  12. 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


Onion | Wheat Germ | Lima Bean Bacteria | Thymus | Yeast | Data Sheet and Questions | Design Your Own DNA Extraction Protocol | Teacher Information | References




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