Gene Expression Analysis by qRT-PCR

Cells and tissues were homogenized and lysed using TRIzol Reagent (Invitrogen) and RNA was extracted using RNeasy Plus Mini kit columns (Qiagen, Hilden, Germany). The cDNAs were synthesized using the GoScriptTM reverse transcriptase kit (Promega, Madison) and assayed using the SYBR® Premix Ex Taq™ II (Takara, Shiga, Japan) and gene‐specific primers listed in supplementary information Table 1 and our earlier study.⁴⁴ (link) Gene expression was determined by the ^2−ΔΔCt method with amplification efficiencies ranging between 90 and 110% for the target and reference genes.

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Matsumae G., Shimizu T., Tian Y., Takahashi D., Ebata T., Alhasan H., Yokota S., Kadoya K., Terkawi M.A, & Iwasaki N. (2021). Targeting thymidine phosphorylase as a potential therapy for bone loss associated with periprosthetic osteolysis. Bioengineering & Translational Medicine, 6(3), e10232.

Publication 2021

TRIzol Reagent RNeasy Plus Mini kit columns GoScriptTM reverse transcriptase kit SYBR® Premix Ex Taq™ II

Cdnas Cells Gene expression Genes Primers Promega Reverse transcriptase Tissues Trizol

Corresponding Organization : Hokkaido University

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Variable analysis

independent variables

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dependent variables

Gene expression

control variables

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controls

Positive control: None mentioned
Negative control: None mentioned

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