Comparison of current methods for total RNA isolation from the small cell tissue cartilage: quality control using capillary electrophoresis. Total RNA was isolated from chondrocytes (cells) and cartilage explants. Cartilage homogenization was performed with scalpel (SC), rotor-stator (RS) or microdismembrator (MD). Different extraction procedures (TRIzol®, RNeasy™, TRIzol®/RNeasy™ and RNAqueous™) were performed as described in the Methods section. Integrity of RNA isolated from different species (human, adult bovine cartilage - cow, and immature bovine cartilage - calf) was analyzed by capillary electrophoresis. 18S and 28S rRNA bands correspond to 41-43 and 47-50 [s], respectively. The RNA yield is specified for each sample [in ng/μl]. After precipitation and washing the RNA was resuspended in different volumes of RNase-free water: 30 μl (Trizol®), 20 μl (RNeasy™), 10 - 20 μl (Trizol®/RNeasy™) and 10 - 20 μl (RNAqueous™). On account of this, the results provided here are comparable with the results provided in Table 1 (μg RNA per 100 mg cartilage). The results are shown by gel electrophoresis (RIN: RNA integrity number; N/A: not available).
Ruettger et al. BMC Research Notes 2010 3:7 doi:10.1186/1756-0500-3-7