Total RNA extraction from Cartilage tumors
Instrument: Precellys® 24 Dual
Sample: Chondrosarcoma
Protocol:
- Precellys kits: 03961-1-007 (CK28R).
- Sample: 50 to 100 mg cartilage tumors (chondrosarcoma).
- Buffer: 1 ml TRIzol Reagent (Invitrogen) + 200 μl Guanidine thiocyanate 4M (SIGMA).
- miRNeasy Mini kit (Qiagen).
- Run the Precellys at 6500 rpm, 3x15sec, 10s break.
Results:
Analysis was performed with subsequent RNA extraction and purification. Verification of RNA quality was performed with an Agilent 2100 Bioanalyzer (Agilent Technologies). The Precellys Dual is successfully validated for total RNA extraction from cartilage tumors. Providing better homogeneity in grinding of hard tissue as cartilage and increase in quality and quantity of total RNA extracted.
Qualification of RNA and miRNA is based on some criteria as 28s/18s ratio, RIN and Agilent 2100 Bioanalyzer profiles. Figures 1 and 2 show the results of RNA and miRNA qualifications extracted from a cartilage tumor. Characteristics of total RNA are reported in Table 1.
RNA and miRNA quality and integrity are optimal to microarray and sequencing analysis.
