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2.7 RNA isolation, reverse transcription and real-time PCR

SD Satya Srirama Karthik Divvela
EO Eric Bekoe Offei
FS Florian Suerland
DG David Revuelta García
JK Julia Kwiatkowski
AB Ajeesh Balakrishnan-Renuka
PB Pauline Bohne
MB Marion Böing
GM Gabriela Morosan-Puopolo
MM Melanie D. Mark
BB Beate Brand-Saberi
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RNA was isolated using TRI reagent (Sigma-Aldrich). 1 µg of RNA was used to perform a reverse transcriptase reaction using GoScript Reverse transcriptase (Promega, Mannheim, Germany). GoTaq qPCR master mix (Promega) was used for Real-time quantitative PCR reaction. These reactions were performed by following the respective manufacturer’s instructions. The Livak method (Livak and Schmittgen, 2001) was used to calculate relative quantification. The list of Real-Time qPCR primers used in the study are available in the supplementary information.

Copyright and License information:  ©2022 Divvela, Offei, Suerland, Revuelta García, Kwiatkowski, Balakrishnan-Renuka, Bohne, Böing, Morosan-Puopolo, Mark and Brand-Saberi.
This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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