Dissecting for RNA preps
From Ucsbgalaxy
Dissection of Ostracod bodies/eyes for RNA extraction:[edit]
This protocol has been used successfully in the field. RNALater could work, but we've had problems retrieving the dissected eyes after preservation. RNALater preserved animals cannot be dissected later as the morphology breaks down.
- keep Trizol on ice
- wear gloves at all times
- Place ostracod in a drop of clean-ish seawater
- Remove carapace and gut
- Transfer the remainder of the animal (basically the “head” ) to a clean drop of seawater
- Dissect median, compound eyes, place in Trizol immediately
- If an eye breaks, make sure that the body material doesn’t get eye fragments
- The easiest way (I have found) to pick up the eyes is to use the 20 uL pipette and suck up a tiny bit of water with the eye. This is why it is important to keep the seawater relatively clean.
- Make sure to use a clean pestle to homogenize the tissue after every 3-5 new eyes/bodies are added to the tube. Check with the marine lab as to how to dispose of phenol-laden homogenizers…
Place samples in Trizol and put tubes in a freezer until they get mailed!
