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Quick Genotyping DNA Preparation Protocol (Zebrafish)

(Please wear protective gloves)
  1. Place zebrafish tissue into PCR tubes, 96-well PCR plate or microtubes. The paraformaldehyde-fixed embryos can also be used.

  2. Add 1 volume Lysis Solution (see Table below) to each sample. Heat to 95°C for 10 – 20 min, then cool to 4°C. Proceed to next step immediately. Use thermo-cycler or heat-block. Typically, 10 min will be sufficient for embryos and 20 min for paraformaldehyde-fixed embryos and adult tissues. The undissolved tissue does not interfere with PCR.

  3. Add 1/10 volume DNA Stabilizing Buffer to each sample and mix. The sample is centrifuged to pellet the debris. 1-5 µl of the supernatant is directly used for 25 µl PCR reaction.

 

Zebrafish tissue
Lysis solution
DNA stabilizing solution
Sperm
50 µl
5 µl
Tail
100 µl
10 µl
Single embryo/larvae
20-100 µl
2-10 µl
Eggs, multiple embryos/larvae
100-500 µl
10-50 µl
 
Note:
  • DNA samples should be stored at 4oC (3 month) or -20oC (> 3 months)
  • The DNA prepared here could not be used for Southern blot analysis.

 

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