【摘要】 This study establishes and validates a method for the precise quantification of aquatic microbial loads using microbial diversity absolute quantitative sequencing. By adding synthetic spike-in DNA to water samples from the Dahei River prior to DNA extraction and 16S rRNA gene sequencing, it generates standard curves to convert sequencing data into absolute microbial copy numbers. The method, which is proved highly accurate(R~2 > 0.99), reveals a clear contrast between the river sites: the upstream community has not only a significantly higher total microbial load but also a completely different makeup of species compared to the downstream site. This approach effectively overcomes the limitations of relative abundance analysis, providing a powerful tool for environmental monitoring, and proposes key steps for future standardization to ensure data comparability and integration.更多还原

【Abstract】 This study establishes and validates a method for the precise quantification of aquatic microbial loads using microbial diversity absolute quantitative sequencing. By adding synthetic spike-in DNA to water samples from the Dahei River prior to DNA extraction and 16S rRNA gene sequencing, it generates standard curves to convert sequencing data into absolute microbial copy numbers. The method, which is proved highly accurate(R~2 > 0.99), reveals a clear contrast between the river sites: the upstream community has not only a significantly higher total microbial load but also a completely different makeup of species compared to the downstream site. This approach effectively overcomes the limitations of relative abundance analysis, providing a powerful tool for environmental monitoring, and proposes key steps for future standardization to ensure data comparability and integration.更多还原