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).
