RNA extraction from subcutaneous adipose tissue (scAT) in human obesit
Instrument: Precellys® 24 Dual
Sample: Subcutaneous adipose tissue (scAT)
Protocol:
- Precellys kits: 03961-1-009 (CKmix).
- Sample: 10 human scAT samples (about 100 to 400 mg/sample) obtained by needle aspiration.
- Buffer: RLT+β-mercaptoethanol.
- Run the Precellys at 6000 rpm, 2x15sec, 20s break.
Results:
Analysis was performed after total RNA extraction using the RNeasy total RNA Mini kit (Qiagen). Total RNA concentration confirmed using NanoDrop ND-1000 and quality using the Agilent 2100 Bioanalyzer (Agilent Technologies). Total yield of RNA was higher with Precellys 24 than manual technique while the purity of RNA remained comparable. Moreover, an amplification factor of the specific gene studied was observed after Precellys 24 homogenization.
Extracted RNA yield obtained with Ultraturax and Precellys methods.
Manual Sample –RIN 7.0; Precellys sample RIN 7.
