* The notes I have say that TdT comes with 5x buffer and CoCl2. But this is an old protocol, so I am not sure if this is still the case... they were purchased from Boehringer Mannheim, but BM was bought out by someone (at least once) and I am not sure what company that is now.
**Fl-dUTP is 2 parts 1mM dUTP: 1 part 1mM FITC-dUTP (very important to dilute the labeled nucleotides).
Tris-Triton buffer is 100mM Tris pH 7.4, 1% Triton X-100, 1 mM EDTA
PBST is 1x PBS + 0.5% Triton X-100.
1) Male 20% paraformaldehyde. Spin down to make sure junk is removed.
2) Wash worms off plate and hypochlorite treat. After larvae and adults are dissolved, wash 3x in water. Meanwhile, make fix mix (above).
3) After 3rd wash, suspend worms in 400ul of water. Chill 5 minutes on ice.
4) Meanwhile, mix together embryo fix (above) and vortex to mix.
5) Add 400ul embryos to tube, invert to mix, immediately freeze in liquid nitrogen.
Protocl uses a swinging eppendorf microfuge to better remove all the supe for each wash. This is not necessary.
Other labeled nucleotides work (rhodamine and DIG) but were not as well tested.