Comparison of different methods for preparation and characterization of total RNA from cartilage samples to uncover osteoarthritis in vivo

Table 1 Variation of Ct-values of specific cartilage genes due to using different RNA isolation methods.

One human cartilage sample was obtained from an OA patient (69 years old) undergoing total knee replacement surgery. Sterile cuts (4 × 50 mg tissue) were prepared and stored in liquid nitrogen. The cartilage samples were reduced to powder using a microdismembrator. After that RNA was isolated according to the respective protocol. After isolation, 0.2 μg RNA was reversed transcribed into cDNA. 1 μl of the cDNA was used as template to amplify human collagen type II messages. QRT-PCR was carried out using a MyiQ Cycler with the iQ SYBR Green Supermix, as described by the manufacturer (Bio-Rad).

ISSN: 1756-0500