SYBR Premix Ex Taq (Perfect Real Time) kit and RT-PCR reagents (Takara Bio) were used for quantification of differentially expressed gene sequences.
The miRNA-125b kit, SYBR Premix Ex Taq™ Perfect Real Time Kit, was purchased from TaKaRa.
First-stranded cDNAs were synthesized using PrimeScript RT Master Mix Perfect Real Time Kit (TaKaRa, China).
The resultant cDNA was amplified by SYBR® Premix Ex Taq™ Perfect Real Time Kit (Takara BIO, Otsu, Japan).
For qPCR analysis, DNA was amplified using SYBR Premix ExTaq Perfect Real Time Kit (Takara Bio Inc. Otsu, Shiga, Japan).
The first-strand cDNA was synthesized using 250 ng of the total RNA template using the PrimeScript RT Master Mix Perfect Real Time kit (Takara Co., Otsu, Japan).
QRT-PCR was performed using SYBR Premix Ex Taq™ (Perfect Real Time) Kit (TaKaRa Biomedicals, Tokyo, Japan) on a LightCycler480 machine (Roche Diagnostics, Basel, Switzerland), according to the manufacturer's instructions.
In addition, relative expression levels of FSCN1 mRNA were assessed by using SYBR Premix Ex Taq (Perfect Real Time) kit (TaKaRa, Dalian, China) and normalized to GAPDH mRNA.
First-strand cDNA was synthesized from 1 μg of total RNA using a PrimeScript™ RT reagent Kit with a gDNA Eraser (Perfect Real Time) kit (Takara, Dalian, China) following the manufacturer's protocol.
Total RNAs from different tissues (leaf, pseudostem and root) and stress-treated leaves were reverse transcribed with PrimeScript™ RT Master Mix (Perfect Real Time) kit (Takara, Japan) into cDNAs for qPCR analysis.
