Plasmid RP4 coupled transfer system expose to PVC leachate for conjugative mechanism investigation

The samples in this experiment were derived from the conjugate cells of the PVC leachate treatment group, which were set up with three replicates in section 2.4.1. Each replicate was treated as an independent transcriptome sample for analysis. The RNA from the conjugation system was extracted using the RNA Pure Bacteria Kit (CWBIO, China) according to the manufacturer's instructions. The purity and integrity of the extracted RNA were assessed by agarose gel electrophoresis and analyzed with a spectrophotometer (Galloptech, China). The extracted RNA was transcribed into cDNA using reverse transcription polymerase chain reaction (RT-PCR) with a reverse transcription kit (Takara, Japan). The cDNA samples were then sent to Novogene (Beijing) for sequencing analysis using the PE150 strategy on the Illumina platform for high-throughput sequencing.

本实验所用样本取自PVC渗滤液处理组的接合细胞，该实验组在2.4.1节中设置了三个生物学重复。每个重复样本均作为独立转录组样本开展分析。采用RNA Pure Bacteria试剂盒（CWBIO，中国），依照产品说明书从接合体系中提取总RNA。通过琼脂糖凝胶电泳对提取RNA的纯度与完整性进行评估，并利用分光光度计（Galloptech，中国）完成检测分析。使用反转录试剂盒（Takara，日本），借助反转录聚合酶链式反应（RT-PCR）将提取的RNA反转录为cDNA。随后将cDNA样本送至北京诺禾致源（Novogene），依托Illumina平台采用PE150测序策略开展高通量测序分析。