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Hot Shot Method of DNA Extraction


Flash content link: https://vlab.amrita.edu/repo/BIOTECH/MOL/HOT_SHOT_METHOD_DNA_EXTRACTION/index.swf




  •   Fins of Zebra fish
  •   Micropipette and pipette tips
  •   Scalpel
  •   Micro centrifuge tubes
  •   Mortar and pestle
  •   Centrifuge
  •   Water bath
  •   4oc freezer
  •  -20oc freezer




  •  50mM NaOH solution
  •  1M Tris-Cl




  •  Pluck out 20-100 mg of fish fin using a sterile scalpel and a forceps.
  •  Homogenize the fins using sterile mortar and pestle.
  •  Aliquot 100 μl of 50 mM NaOH in to a microfuge tube.
  • Transfer the homogenized fins of zebra fish into this micro centrifuge tube.
  •  Incubate at 95°C for 20min in a water bath.
  • After the incubation, the tubes are allowed to cool at 4°C.
  •  Add 1/10th volume of 1 M Tris-HCl (pH 8.0) for neutralizing the basic solution.
  • Centrifuge the sample at 5000rpm for 10min to pellet the debris. The supernatant obtained is collected in afresh micro centrifuge tube  (1-5μl of this supernatant can be used immediately  per 25 μL of  PCR reaction mixture).
  • Store the sample for at least 3 months at 4°C or longer at -20°C freezer.





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