RNA Extraction from Human Skin Biopsies

Instrument: Precellys® 24 Dual

Sample:  Human skin biopsies

Protocol:

  • Precellys kits: CK14 (small ceramic beads)
  • Sample : human skin biopsies (40-70mg)
  • Buffer : RA1 lysis buffer (Macherey- Nagel) +1% ß- Mercapto ethanol (350μl)
  • Run the Precellys®24 at 6300 rpm, 6x23 sec., 2 min. break (Samples are chilled on ice between each cycle) Cryolys Option should be used to keep the samples at low temperature.

Results:

Compared to classical lysis (chaotropics ions + ß- Mercapto ethanol + Vortex), RNA yield increased from 2 to 4 fold (Nanodrop quatification). RNA quality was assessed using a 2100 Bioanalyser. RNA Integrity Number (RIN) is over 8, meaning good quality.

 

RNA extracted in highest quantity using Precellys®24 allowed us to study transcription modifications of several genes implicated in pigmentation or inflammation after UV-solarspectrum-radiation corresponding to 1 to 2 MED (Minimal Erythema Dose).