Hello community! I’m embarking on a RNA seq study and I’m wondering if anyone has some advice on RNA isolation. I’m wondering about the number of liquid nitrogen freeze thaw cycles you’ve used and I have some other pretty simple questions. Thanks!
six cycles of freeze thaw between a dry ice/ethanol bath and a 37C water bath work well for our RNA preps.
A lot depends on what you’re doing and the quantity of worms. For very large amounts of worms - pellets in the milliliters - I grind them using a mortar and pestle, keeping them cold the entire time by adding liquid nitrogen extremely often. For more moderate numbers of worms or for very small numbers I use proteinase K, and a single freeze/thaw. I know other methods recommend repeated freeze/thaw cycles, but I haven’t used those methods.
Thanks so much - we will incorporate these suggestions