Table 1

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

Ct-values (mRNA-Expression)


METHOD

Col II

(109 bp)

Col II

(609 bp)

GAPDH

(254 bp)

ΔCt1

ΔCt2


Trizol®

23.1

28.9

23.5

-0.4

5.4

RNeasy™

23.2

28.5

23,7

-0.5

4.8

Trizol®/RNeasy™

23.2

26.8

23.8

-0.6

3.0

RNAqueous

24.1

24.5

23.5

0.6

1.0


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).

Ruettger et al. BMC Research Notes 2010 3:7   doi:10.1186/1756-0500-3-7

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