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

JS Jing Shi
YQ Yiping Qu
XL Xinru Li
FS Fang Sui
DY Demao Yao
QY Qi Yang
BS Bingyin Shi
MJ Meiju Ji
PH Peng Hou
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Total RNA from tissues and cell lines were extracted using Trizol reagent (Takara Inc., Dalian, P.R. China) following the manufacturer's protocol. The cDNA was synthesized with 500 ng total RNA by using PrimeScript RT reagent Kit (Takara Inc., Dalian, P.R. China). Quantitative RT-PCR (qRT-PCR) was carried out on a CFX96 Thermal Cycler DiceTM real-time PCR system (Bio-Rad Laboratories, Inc., CA) using SYBR Premix Ex TaqTM (Takara Inc., Dalian, P.R. China). The mRNA expression of the indicated genes was normalized to 18S rRNA cDNA. Each sample was run in triplicate. The primer sequences were presented in Supplementary Table 4.

Copyright and License information:  ©2016 The Author(s)
Cell Death and Disease is an open-access journal published by Nature Publishing Group. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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