Quantitative gene expression analysis

Total RNA was extracted with the RNA Extraction kit (Beyotime, Shanghai, China), and the concentrations were measured with the NanoDrop 2000 (Thermo Fisher Scientific, MA, USA). The cDNA was synthesized using the RevertAid Master Mix (Thermo Fisher Scientific). Real-time quantitative polymerase chain reaction (RT-qPCR; Platinum® qPCR SuperMix-UDG, Thermo Fisher Scientific) was performed by a CFX96 Touch qPCR System (Bio-Rad, CA, USA) as previously described [23 (link)], using specific sets of primers/probes for genes of interest (Supplementary Table 1). Gene expressions were assessed in relation to the levels of β-actin.

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Liang J., Chen B., Li Y., Nie D., Lei C., Yang Q., Zhou Y., Li S., Chen S., Li B., Shu L., Chen L, & Wang W. (2022). Asymptomatic Hyperuricemia Is Associated with Achilles Tendon Rupture through Disrupting the Normal Functions of Tendon Stem/Progenitor Cells. Stem Cells International, 2022, 6795573.

Publication 2022

RNA Extraction kit NanoDrop 2000 RevertAid Master Mix CFX96 Touch qPCR System

Actin Cdna Gene expressions Genes Platinum Primers Real time polymerase chain reaction Touch

Corresponding Organization : Wuhan University

Other organizations : Xiangnan University, Huazhong University of Science and Technology, Tongji Hospital, Shanghai Changzheng Hospital

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Gene expressions

control variables

β-actin gene expression levels

controls

Positive control: None mentioned
Negative control: None mentioned

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