Isolating Complete Microbial Genomes from Small and Heterogeneous Samples

完整基因组序列的测定依赖于纯化相对大量的细胞或DNA的能力。但是，由于大多数微生物物种不能被培养或与其他生物体紧密结合生活，因此难以获得足够用于产生这些生物体的全基因组序列的大的且未污染的细胞样品。拟议研究的目的是开发用于从小细胞和/或混合细胞群体中回收和纯化足够质量和数量的DNA以进行基因组测序的方法。使用细胞和DNA富集程序的组合，沿着全基因组扩增方法，该项目的最终目标是设计有利于从单个细胞分析完整微生物基因组的条件。

The determination of complete genomic sequences relies upon the ability to purify relatively large quantities of cells or DNA. But because the majority of microbial species can not be cultivated or live in close association with other organisms, it is difficult to obtain large and uncontaminated samples of cells sufficient for producing whole genome sequences of these organisms. The aims of the proposed research are to develop methods for recovering and purifying DNA of sufficient quality and quantity for genomic sequencing from small and/or mixed populations of cells. Using a combination of cell and DNA enrichment procedures, along with whole genome amplification methods, the ultimate goal of this project is to devise conditions that would facilitate the analysis of a complete microbial genome from a single cell.