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Total RNA was extracted using Trizol (Thermo Fisher Scientific) according to the method previously described [26]. For mRNA relative expression analysis, total RNA was reversed transcribed into cDNA with Prime-Script RT kit (Takara, Shiga, Japan) and amplified with SYBR-Green Real-time PCR Master Mix (Applied Biosystems, Thermo Fisher Scientific). The mRNA expression level of GAPDH was used as an internal normalization control. Comparative quantification was performed by using the 2-ΔΔCt method. All primers are available in the Additional file 1: Table S1.
Copyright and License information: The Author(s). ©2017
Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
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