Table 2.

The Conditions of Each Step of RT-qPCR

Methods	Conditions
System of reverse transcription (Reverse Aid First Strand cDNA Synthesis Kit)	2 μL 0.1 ng - 5 μg total RNA
	1 μL Oligo (dT)18 primer
	9 μL Water, nuclease-free
	4 μL 5× Reaction Buffer
	1 μL RiboLock RNase Inhibitor (20 U/μL)
	2 μL 10 mM dNTP Mix
	1 μL RevertAid M-MuLV RT (200 U/μL)
	Total volume 20 μL
Conditions of reverse transcription	Incubate for 60 min at 42°C → Terminate the reaction by heating at 70°C for 5 min
System of RT-PCR	10 μL SYBR Green PCR Master Mix
	1 μL Upstream primer
	1 μL Downstream primer
	1 μL cDNA template
	7 μL Diethyl pyrocarbonate
	Total volume 20 μL
Cycle conditions of RT-qPCR	Pre-denaturation at 95°C for 3 min → denaturation at 95°C for 15 s, annealing at 60°C for 30 s, and extension at 72°C for 15 s, 40 cycles → dissolution curve was completed at 95°C for 30 s, 60°C for 30 s, and 95°C for 15 s