I want to do RNA extraction for zebrafish embryos/larvae at 2dpf, 5dpf and 7dpf. I'm using Trizol and I want to store the larvae in Trizol at -20 oC first before extraction at a later time. However, the protocol I found so far are for larvae younger than 5dpf, and they are homogenize in Trizol before stored. None included euthanasia. At 7dpf, the zebrafish can swim around, so I feel the need to euthanize them in Benzocaine, but I'm afraid if they will affect RNA extraction in anyway.
After the animal has died, RNA degradation can begin to happen quickly (within 1-2 minutes) because cells deprived of oxygen begin to release endogenous RNases outside of the cellular compartments where they belong. To maximise the RNA quality it is important that RNA is protected from RNases (either by snap-freezing or homogenisation in chaotropic lysis reagent like TRIzol) as quickly as possible after the animal's death. Any method of euthanasia which causes tissue to be deprived of oxygen for a significant amount of time (eg long period of unconsciousness with reduced respiration/circulation precedes death) would have the potential to reduce RNA quality, but if the euthanasia method is rapid this shouldn't have any effect on your RNA. I don't know anything about benzocaine but when I briefly worked in a zebrafish lab we used ice water for euthanasia and it was basically instantaneous, perhaps this would work for you also.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA