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Real-time quantitative reverse transcription PCR (qRT-PCR)

HH Haiyan Huang
WY Wenjun Yu
RW Rongshui Wang
HL Huili Li
HX Huijun Xie
SW Shuning Wang
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To confirm the results of RNA sequencing, five genes (ndhA, ndhB, hno, pno and hsh) were chosen for qRT-PCR analysis. Cells were harvested at the early-exponential phase and total RNA was extracted using Ezol Total RNA Extraction Reagent (Shanghai GenePharma, China) according to the manufacturer’s protocol. During this process, DNA was removed by digestion with DNaseI. Total cDNA was synthesized with TransScript First-Strand cDNA Synthesis SuperMix (Beijing TransGen Biotech, China). qPCR was performed by using SYBR Real-Time PCR Master Mix (Shanghai GenePharma, China) according to the manufacturer’s protocol. The 16S rRNA gene was used as the reference gene. Reactions were performed in triplicate. The primers employed in amplification are listed in the Supplementary Table S1.

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