miRNA qPCR Reaction Protocol

MiRNA qPCR reactions were performed using the TaqMan MicroRNA reverse transcription kit (Life Technologies), Taqman MicroRNA qPCR assays (Life Technologies) and TaqMan Universal PCR Master Mix, as previously described [25 (link), 26 (link)]. Briefly, cDNA was reverse transcribed from total RNA using specific miRNA primers according to the suggested reaction conditions on a BioRad T100 thermal cycler. Quantitative polymerase chain reaction (qPCR) was performed using a ViiA 7 Real Time PCR System (Applied Biosystems, Carlsbad, CA). Sno202 was used as a housekeeping control RNA. Relative expression was calculated using the 2^−ΔΔCtmethod.

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Zhou H., Zhang J., Eyers F., Xiang Y., Herbert C., Tay H.L., Foster P.S, & Yang M. (2016). Identification of the microRNA networks contributing to macrophage differentiation and function. Oncotarget, 7(20), 28806-28820.

Publication 2016

TaqMan MicroRNA reverse transcription kit Taqman MicroRNA qPCR assays TaqMan Universal PCR Master Mix T100 thermal cycler ViiA 7 Real Time PCR System

Assays Cdna Mirna Primers Reverse transcription

Corresponding Organization :

Other organizations : Second Affiliated Hospital of Jilin University, Hunter Medical Research Institute, Central South University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Relative expression of miRNA

control variables

Sno202 (used as a housekeeping control RNA)

controls

Positive control: None mentioned
Negative control: None mentioned

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