期刊
ISME JOURNAL
卷 4, 期 10, 页码 1352-1356出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/ismej.2010.73
关键词
fluorescence in situ hybridization (FISH); fluorescence-activated cell sorting (FACS); cell fixation; genomics
资金
- Office of Science of the US Department of Energy [DE-AC02-05CH11231]
We modified the standard ribosomal RNA-targeted fluorescence in situ hybridization (FISH) protocol by removing the fixation steps to allow recovery of unmodified nucleic acids. Using this method, hybridized cells could be visualized in two bioreactor sludges and termite hindgut samples by epifluorescence microscopy. We then targeted one bacterial and one archaeal population in the sludge samples with group-specific oligonucleotide probes using in-solution fixation-free FISH and sorted hybridized populations using fluorescence-activated cell sorting (FACS). We could show that sorted populations were highly enriched for the target organisms based on 16S rRNA gene sequencing, thus confirming probe specificity using the modified FISH protocol. This approach should facilitate subsequent genomic sequencing and analysis of targeted populations as DNA is not compromised by crosslinking during fixation. The ISME Journal (2010) 4, 1352-1356; doi: 10.1038/ismej.2010.73; published online 27 May 2010
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据