INTRO
Continued RNA isolation for this project, which was initially started on 20250804.
I isolated RNA from the remaining 100 tissue samples. See RNA quantification on 20250910 for the full list of samples.
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 containing ~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.
RNA Isolation
RNA was isolated using the Quick-DNA/RNA MiniPrep Plus Kit, according to the manufacturer’s protocol with the following changes/notes:
Treated with optional DNase step.
Eluted with 50uL of H2O.
Samples were stored in Valentina’s original boxes in the middle -80oC for quantification at a later date.
The following seven samples yielded no RNA when they were quantified on 20250910 and their tubes were discarded:
C4PCC5PCC1PMC2PMC3PMC4PMC5PM