Sybr premix ex taq 2 with rox

Total RNA was extracted from prostate cancer cells and normal cells with standard TRIzol^® solution (Invitrogen; Thermo Fisher Scientific, Inc.). For miRNA expression, RT reactions were performed with a One Step PrimeScript miRNA cDNA Synthesis kit (Takara Biotechnology Co., Ltd.), followed by PCR with SYBR^® Premix Ex Taq (Takara Biotechnology Co., Ltd). For mRNA, cDNA was synthesized from the total RNA using a PrimeScript RT Reagent Kit (Takara Biotechnology Co., Ltd.). qPCR amplification reactions for CCND1 expression were performed with SYBR^® Premix Ex Taq II with ROX (Takara Biotechnology Co., Ltd.,). For miRNA and mRNA amplifications, analysis was performed with the ABI 7500 Fast Real-Time PCR system (Applied Biosystems; Thermo Fisher Scientific, Inc.). According to supplier's instructions, the PCR conditions consisted of 95°C for 30 sec, followed by 40 cycles of amplification (95°C for 3 sec and 60°C for 30 sec). All fold changes were calculated using the comparative Cq (ΔΔCq) method using GAPDH for normalization (13 (link)). All relevant primer sequences are listed in Table I, and details of the reaction mixtures for RT and qPCR are listed in Tables II and III.

Total RNA was extracted from prostate cancer cells and normal cells with standard TRIzol^® solution (Invitrogen; Thermo Fisher Scientific, Inc.). For miRNA expression, RT reactions were performed with a One Step PrimeScript miRNA cDNA Synthesis kit (Takara Biotechnology Co., Ltd.), followed by PCR with SYBR^® Premix Ex Taq (Takara Biotechnology Co., Ltd.). For mRNA, cDNA was synthesized from the total RNA using a PrimeScript RT Reagent Kit (Takara Biotechnology Co., Ltd.). qPCR amplification reactions were performed with SYBR^® Premix Ex Taq II with ROX (Takara Biotechnology Co., Ltd.).