摘要:土壤宏转录组学是通过制备土壤 RNA 样本、RNA 测序、以及利用一系列生物信 息学方法和平台搭建来完成土壤微生物组的转录过程分析,提供关于基因表达和土壤微 生物组功能活性,从而获得微生物组关键代谢差异表征等信息的一门学科。关键点是针 对土壤 RNA 样本,表征特定条件下执行各个代谢过程的微生物活性特征,极大地规避 了因高通量 DNA 测序带来无法准确反映土壤微生物代谢活性的缺陷。本实验以两种类 型(酸性和碱性)湿地土壤样本为例,详述了利用市售 RNA 提取试剂盒进行的土壤宏 转录组样本制备流程,为准确评价土壤 RNA 样本制备提供参考,同时给出了宏转录组 数据分析流程,为从 RNA 水平分析土壤微生物表达活性提供思路。 关键词:土壤,宏转录组学,RNA,土壤微生物代谢活性 材料与试剂 1. 50 ml 离心管 (Thermo Fisher Scientific, USA) 2. 酚氯仿异戊醇 (配比 25:24:1, pH=8) 3. 琼脂糖 4. 电泳缓冲液 (TAE) 5. RNA 提取试剂盒 RNA Mini Kit (Qiagen, Hilden, Germany)
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