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
- Micro centrifuge tubes
- Mortar and pestle
- 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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