Real-Time PCR for Gene Expression

Total RNA was isolated with Trizol reagent and synthesized cDNA using the superscript III preamplification system (Invitrogen). The real-time polymerase chain reaction (PCR) was performed using a CFX Connect Real-Time PCR system (Bio-Rad Laboratories, Foster city, CA, USA), as previously described [29 (link)]. FAM dye-labeled TaqMan MGB probes and PCR primers for human IL-6 (Hs00985639_m1) and HO-1 ((Hs00157965_m1) were purchased from Applied Biosystems (Foster City, CA, USA). For the internal positive control, β-Actin (Hs01060665_g1) was used with a FAM dye-labeled TaqMan MGB probe.

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Chiang K.C., Chang K.S., Hsu S.Y., Sung H.C., Feng T.H., Chao M, & Juang H.H. (2020). Human Heme Oxygenase-1 Induced by Interleukin-6 via JAK/STAT3 Pathways Is a Tumor Suppressor Gene in Hepatoma Cells. Antioxidants, 9(3), 251.

Publication 2020

superscript III preamplification system CFX Connect Real-Time PCR system

Actin Cdna Human Polymerase chain reaction Primers Real time polymerase chain reaction Trizol

Corresponding Organization : Chang Gung Memorial Hospital

Other organizations : Min Sheng General Hospital

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Variable analysis

independent variables

Not explicitly mentioned

dependent variables

Expression of IL-6
Expression of HO-1

control variables

Expression of β-Actin

controls

Positive control: β-Actin (Hs01060665_g1) with FAM dye-labeled TaqMan MGB probe

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