RNA Extraction and qRT-PCR Analysis in Cell Samples

After extracting the RNA from adherent cells, exfoliated cells and culture medium were collected for RNA extraction by TRIZOL regent as we previously reported [15 (link)], and cDNAs were synthesized with PrimeScript™ IV 1st strand cDNA Synthesis Mix (Takara, Beijing, China) for mRNA and Mir-X™ miRNA First-Strand Synthesis (Takara Bio USA, Inc., San Jose, CA, USA) for miRNA. QRT-PCR analysis was measured using the Taq Pro Universal SYBR qPCR Master Mix (Vazyme Biotech Co., Ltd., Nanjing, China). Each assay was carried out in triplicate by the Light Cycler 480 Instrument (Roche, Basel, Switzerland). The primers for RT-qPCR are provided in the Appendix B (Table A4).

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Li Y., Li K., Lou X., Wu Y., Seery S., Xu D., Pei Y., Qian B., Wu Y., Liang S., Wu K, & Cui W. (2023). HNRNPA2B1-Mediated MicroRNA-92a Upregulation and Section Acts as a Promising Noninvasive Diagnostic Biomarker in Colorectal Cancer. Cancers, 15(4), 1367.

Publication 2023

PrimeScript™ IV 1st strand cDNA Synthesis Mix Mir-X™ miRNA First-Strand Synthesis Taq Pro Universal SYBR qPCR Master Mix Light Cycler 480 Instrument

Assay Cdna Cells Culture medium Light Mirna Mrna Primers Synthesis Trizol

Corresponding Organization : Wenzhou Medical University

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Variable analysis

independent variables

RNA extraction method (TRIZOL reagent)

dependent variables

MRNA expression levels
MiRNA expression levels

control variables

Extraction of RNA from adherent cells, exfoliated cells, and culture medium
CDNA synthesis for mRNA and miRNA using specific kits
QRT-PCR analysis using specific master mix
Triplicate measurements using Light Cycler 480 Instrument

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