Quantitative Analysis of miRNA and mRNA

RNA extraction and quantitative RT-PCR (qRT-PCR) were carried out as we previously described [26 (link),27 (link),28 (link)]. The differential miRNA expression patterns were validated with qRT-PCR using a TaqMan assay (Applied Biosystems, Waltham, MA, USA), or NCode VILO miRNA cDNA Synthesis kit and EXPRESS SYBR GreenER miRNA qRT-PCR kit (Invitrogen, Carlsbad, CA, USA). The mRNA levels were assessed with qRT-PCR using SYBR Green dye (Applied Biosystems). RNU48 (for TaqMan qRT-PCR) or 5.8S (for SYBR qRT-PCR), and GAPDH were used as endogenous controls of miRNA and mRNA qRT-PCR, respectively. Each sample was run in triplicate. The primers used are listed in Table S2.

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Mo J.S., Park W.C., Choi S.C., Yun K.J, & Chae S.C. (2019). MicroRNA 452 Regulates Cell Proliferation, Cell Migration, and Angiogenesis in Colorectal Cancer by Suppressing VEGFA Expression. Cancers, 11(10), 1613.

Publication 2019

TaqMan assay NCode VILO miRNA cDNA Synthesis kit EXPRESS SYBR GreenER miRNA qRT-PCR kit SYBR Green dye

Assay Cdna Gapdh Mirna Mrna Primers Rt pcr Sybr green Synthesis

Corresponding Organization : Wonkwang University

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Variable analysis

independent variables

RNA extraction and quantitative RT-PCR (qRT-PCR) protocols

dependent variables

Differential miRNA expression patterns
MRNA levels

control variables

RNU48 (for TaqMan qRT-PCR) or 5.8S (for SYBR qRT-PCR) as endogenous controls of miRNA qRT-PCR
GAPDH as endogenous control of mRNA qRT-PCR

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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