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RNA extraction from torafugu pufferfish

Instrument: Minilys homogenizer

Instrument: Precellys® 24 Dual

Sample:  Torafugu pufferfish (Takifugu rubripes) liver or muscle

Protocol:

  • Precellys kits: 03961-1-002 (2.8 mm ceramic beads).
  • Add Sample: 100 mg of liver or muscle cut into 7 mm cubic pieces of torafugu pufferfish (Takifugu rubripes 33cm, 750g) stored in RNAlater at -20°C.
  • Add Buffer: 1 mL Lysis reagent of Qiagen RNeasy lipid tissue mini kit.
  • Run the Minilys at 5000 or 4000 rpm, 2x30sec (15sec break at RT).
  • Run the Precellys at 5000 rpm, 2x30sec (15sec break at RT).

Results:

Analysis performed on denatured agarose gel (0.9%) electrophoresis stained with Sybr Green. The amount of RNA was determined using a spectrophotometer. The quality of extracted RNA from torafugu pufferfish was good. For the torafugu pufferfish muscle, which is exceptionally hard among the fish muscle, 5000 rpm 2x 30sec gave very good RNA yield. The 4000 rpm treatment was not enough (the homogenate looked somewhat granular). The RNA concentration extracted from the fish liver using Minilys was somewhat smaller than the amount of RNA extracted using Precellys24.

concentration of RNA extracted (μg/μl)