Development of single-cell sequencing technology for microbial populations

当在DNA水平（研究基因型）或RNA水平（研究基因表达）分析生物样品时。我们目前必须一次分析数千或数百万个细胞。这是一种非常有用的方法，但这意味着我们错过了生物系统中的许多复杂性，因为在大多数情况下，微生物培养物中的DNA在每个细胞中可能是不同的，每个细胞中的基因表达也是异质的。这对于在表面上形成生物膜的群落尤其重要，因为可能有不同的菌株或细菌物种一起工作，在群落中发挥不同的作用。生物膜对于逃避抗菌剂处理的细菌尤其重要。在这个项目中，我们的目标是开发微生物单细胞测序的方法，并针对许多不同的用例进行测试。我们将使用荧光激活细胞分选（FACS）来分离细胞，然后通过使用分子条形码方法开发并行分析这些细胞的方案。

When biological samples are analysed at the DNA level (to study genotypes) or the RNA level) to look at gene expression. We currently have to analyze many thousands or millions of cells at a time. This has been a very informative approach but it means that we miss a lot of complexity in biological systems, as in most cases the DNA in a microbial culture may be different in each cell and the gene expression in each cell is also hetrogeneous. This is particularly important is communities that form biofilms on surfaces as there may be different strains or species of bacteria working together performing different roles in the community. Biofilms are especially important for bacteria evading antimicrobial treatment. In this project we aim to develop the methodologies for single cell sequencing of microbes and test these against a number of different use cases. We will use Fluorescence Activated Cell Sorting (FACS) to separate cells and then develop protocols to analyze these cells in parallel by using molecular barcoding methods.