INTRO
Steven tasked me with helping and/or isolating RNA (GitHub Issue) for Valentina (visiting scientist from Cristian’s lab in Chile). Samples were whole body (no shells) of small juvenile oysters exposed to poly I:C (Wikipedia) and then subjected to mechanical stress or temperature stress (or no stress at all).
I previously helped isolate RNA from a subset of these on 20250804 and needed to process the remainder, now that Valentina has returned to Chile.
MATERIALS & METHODS
Whole body tissue had been snap frozen and stored at -80oC.
Homogenization
300uL of DNA/RNA Shield (ZymoResearch) were added to the frozen tissue. Samples and DNA/RNA Shield were transferred to 1.5mL SafeLock (Eppendorf) microcentrifuge tubes containig ~50uL of 0.5mm zirconium oxide beads and ~50uL of 0.15mm zirconium oxide beads. Samples were homogenized using the Roberts Lab Bullet Blender 5E Gold + (NextAdvance) with 1.5mL tube adapters for 3mins at speed 12. The Bullet Blender was pre-cooled, and cooled, using dry ice.
Homogenized samples were stored @ -20oC until RNA isolation can be performed with Quick-DNA/RNA MiniPrep Plus Kit (ZymoResearch) later this week.