RNA Isolation and qRT-PCR in RAW264.7 Macrophages

Total RNA was isolated from RAW264.7 macrophages using RNAiso Plus (TaKaRa) [33 (link), 34 (link)]. RNA was converted into cDNA using PrimeScript RT Mix (TaKaRa). Afterward, qRT-PCR experiments were conducted in a LightCycler PCR system (Roche) with UNICONTM SYBR Green (11198ES08, Yeasen). The sequences of all the primers used are shown in Table 1.

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Qiu J., Fu Y., Chen Z., Zhang L., Li L., Liang D., Wei F., Wen Z., Wang Y, & Liang S. (2021). BTK Promotes Atherosclerosis by Regulating Oxidative Stress, Mitochondrial Injury, and ER Stress of Macrophages. Oxidative Medicine and Cellular Longevity, 2021, 9972413.

Publication 2021

RNAiso Plus PrimeScript RT Mix LightCycler PCR system

Cdna Macrophages Primers Sybr green

Corresponding Organization : Sun Yat-sen University

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Variable analysis

independent variables

RNA isolation method (RNAiso Plus)

dependent variables

Gene expression (measured by qRT-PCR)

control variables

Cell type (RAW264.7 macrophages)
CDNA synthesis method (PrimeScript RT Mix)
QRT-PCR system (LightCycler PCR system)
QRT-PCR reagent (UNICONTM SYBR Green)

controls

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