Ferrari Belinda C, Winsley Tristrom J, Bergquist Peter L, Van Dorst Josie
School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Randwick, NSW, Australia.
Methods Mol Biol. 2012;881:3-26. doi: 10.1007/978-1-61779-827-6_1.
The isolation and subsequent characterization of microbial cells from within environmental samples is a difficult process. Flow cytometry and cell sorting, when combined with the application of fluorescent probes, have the capability for the detection and separation of diverse microbial populations from within complex mixtures. The isolation of single cells allows for downstream investigations towards system-level characterization of unknown Bacterial Phyla to occur. We describe here the combination of fluorescent in situ hybridization and cell sorting for the detection and isolation of Candidate Division TM7 bacteria from an enriched soil sample. The result is the isolation of rare cells suitable for advanced molecular analysis including whole genome amplification and high-throughput pyrosequencing.
从环境样本中分离并随后鉴定微生物细胞是一个困难的过程。流式细胞术和细胞分选技术，当与荧光探针结合应用时，有能力从复杂混合物中检测和分离不同的微生物群体。单细胞的分离使得能够对未知细菌门进行系统水平的表征的下游研究得以开展。我们在此描述荧光原位杂交和细胞分选相结合用于从富集土壤样本中检测和分离候选分类单元TM7细菌。结果是分离出了适合进行包括全基因组扩增和高通量焦磷酸测序在内的高级分子分析的稀有细胞。
