Single worm lysis and PCR
- Prefill PCR tubes with 5 µl lysis buffer without proteinase K. Strips of PCR tubes work well.
- Pick single worms to a fresh plate without food. When the worm crawls away from the bit of food transferred with the worm, use a pipette with a 10µl tip to put a drop of 2 µl of lysis buffer on the worm. Then pipette the worm off from the plate and into the prefilled PCR tube.
- When all of the worms have been picked, quick freeze the tube in liquid N2, and thaw.
- Add 5 µl lysis buffer containing proteinase K
- Incubate 1 hour at 60°C to lyse the worm
- Incubate 15 min. at 95°C to inactivate proteinase K
- Use 2 µl in a PCR reaction
|Worm lysis buffer|
|Tris-HCl pH 8.3||10 mM|
Just before use, add 6 µl of 10 mg/ml proteinase K to 0.5 ml of lysis mix.