RNA isolation and quantitative real-time PCR

RF Runping Fang
XC Xin Chen
SZ Sicong Zhang
HS Hui Shi
YY Youqiong Ye
HS Hailing Shi
ZZ Zhongyu Zou
PL Peng Li
QG Qing Guo
LM Li Ma
CH Chuan He
SH Suyun Huang
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For RNA isolation, TRIzol reagent (Life Technologies) was added to the indicated GSC or LN229 cells, and total RNA was extracted from the cells. mRNA was purified from the total RNA by poly-A selection with GenElute™ mRNA Miniprep Kit (Sigma). cDNAs were synthesized from the purified mRNA by using iScriptTM Reverse Transcription Supermix (Bio-Rad). For quantitative real-time PCR, the PCR reactions were set with PowerUpTM SYBR® Green Master Mix (Life Technologies) on a 7500 Fast Real-time PCR System (Applied Biosystems) following the manufacturer’s instruction. Primers used in this study were summarized in Supplementary Table 1.

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