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RT-qPCR validation of microarray unigenes of P. euphratica roots in response to soil drying

AI Arshad Iqbal
TW Tianxiang Wang
GW Guodong Wu
WT Wensi Tang
CZ Chen Zhu
DW Dapeng Wang
YL Yi Li
HW Huafang Wang
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Total root RNA was isolated from all lines (1–5) treated with RNase-free DNase (Chang et al., 2012). The root RNA samples were reverse-transcribed with a cDNA Synthesis Kit (CWBIO Inc., Beijing, China). cDNA products were used for SYBR Green-based RT-qPCR analysis, each sample was run in triplicate. The RT-qPCR running conditions were: 95 °C for 10 min, followed by 40 cycles of 95 °C for 15 s, 52 °C for 20 s, and 72 °C for 30 s, with a final step of 72 °C for 10 min. Using the roots of line 5 group plants as control, the expression levels of differential expression genes (DEGs) were calculated using the 2−∆∆Ct method61.

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