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RNA extraction and quantitative polymerase chain reaction

CC Ching-Yu Chuang
CY Chih-Chao Yang
BS Bing-Wen Soong
CY Chun-Ying Yu
SC Shu-Hwa Chen
HH Hsiang-Po Huang
HK Hung-Chih Kuo
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Total RNA was extracted with RNeasy Mini Kit (Qiagen, Germantown, MD, USA) and treated with DNase I (Qiagen). RNA was converted to cDNA using the High Capacity cDNA Reverse Transcriptase Kit (Life Technologies). Polymerase chain reaction (PCR) was performed using the Platinum Taq DNA polymerase kit (Life Technologies) according to the manufacturer’s instructions. Reverse transcriptase quantitative PCR (RT-qPCR) was performed with the SYBR® Select Master Mix (Life Technologies), and analyzed with the 7900 real-time PCR system (Life Technologies). Gene specific primers are listed in Table S2. Results were normalized to GAPDH, and relative expression was calculated according to the ΔΔCt method.

Copyright and License information: The Author(s) ©2019
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