Quantifying Osteoclast-Related Gene Expression

Total RNA was extracted using the TRIzol reagent (Invitrogen). First-strand cDNA was synthesized with the Superscript III First-Strand Synthesis System (Invitrogen) using oligo dT primers; qRT-PCR was then performed in triplicate on a Light Cycler 480 (Roche, Mannheim, Germany) using Probes Master Mix (Roche) [37 (link)]. Primers for Trap (Mm00475698_m1), CatK (Mm00484039_m1), Ctr (Mm00432282_m1), Mmp9 (Mm00442991_m1), Opg (Mm01205928_m1), and Rankl (Mm00441906_m1) were obtained from Applied Biosystems (Foster City, CA, USA). The threshold cycle (Ct) value for each gene was normalized to the Ct value of 18S rRNA (Hs03928990_g1).

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Lee J.Y., Kim D.A., Kim E.Y., Chang E.J., Park S.J, & Kim B.J. (2021). Lumican Inhibits Osteoclastogenesis and Bone Resorption by Suppressing Akt Activity. International Journal of Molecular Sciences, 22(9), 4717.

Publication 2021

TRIzol reagent Superscript III First-Strand Synthesis System Light Cycler 480 Probes Master Mix

18s rrna Cdna Gene Light Mmp9 Oligo dt Primers Rankl Synthesis Trizol

Corresponding Organization : Asan Medical Center

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Expression levels of Trap, CatK, Ctr, Mmp9, Opg, and Rankl

control variables

18S rRNA expression

controls

Positive control: None mentioned
Negative control: None mentioned

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