Hello. I am doing an RIP protocol that uses UV crosslinking in a stratalinker. I have spun down the worms and plated on LB media (blank plate no food) and set the energy for 3000. I put them ~10cm from the UV lamp with pipette boxes. However, when I take the worms out, they are still moving as vibrant as ever! I expected them to be immobilized (or dead). Is this normal? What could I be doing differently? Thanks!
don’t worry…they slow down in the dry ice-ethanol ethanol bath…
The energy (3kJ/m2) and the distance seem fine to me based upon available CLIP protocols for worms.
Other factors to bear in mind include the age of the worms, younger worms are more sensitive than older worms. Also irradiation causes arrest (e.g. cesation of further growth) but not necessarily instant death.