Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

Quantitative reverse transcriptase PCR (qRT-PCR) analysis

FG Federica Giannetti
PB Patrizia Benzoni
GC Giulia Campostrini
RM Raffaella Milanesi
AB Annalisa Bucchi
MB Mirko Baruscotti
PD Patrizia Dell’Era
AR Alessandra Rossini
AB Andrea Barbuti
ask Ask a question
Favorite

Total RNA was isolated using TRIzol (Thermo Fisher Scientific). GoScript™ Reverse Transcription System (Promega) was used to synthesize cDNA following the manufacturer’s instructions. For each gene, qRT-PCR was performed on technical duplicates or triplicates from at least 4 independent experiments, using 10 ng of cDNA with the iQTMSYBR® Green Supermix (Bio-Rad) using the iCycler Bioer System (BIOER). Expression data were analysed using 2^(-ΔCT) method using β-actin (ACTB) as housekeeping gene. Gene expression levels were normalized to cardiac Troponin-T (TNNT2) levels to account for differences in cardiomyocytes yield among various differentiation experiments. Primers used are given below.

Copyright and License information: The Author(s) ©2021
Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

post Post a Question
0 Q&A