Linkage between Light Microscopic Observations and Molecular Analysis by Single-Cell PCR for Ciliates

• Shimano Satoshi

  Environment Education Center, Miyagi University of Education

• Sanbe Mitsuo

  Environment Education Center, Miyagi University of Education

• Kasahara Yasuhiro

  Institute of Low Temperature Science, Hokkaido University

Abstract

The molecular analysis of ciliates for identification and phylogeny is usually conducted through PCR amplification and DNA sequencing, with DNA extracted from a large number of cells. Therefore, the analysis of ciliates is limited to only those species that can be cultured. We propose a single-cell PCR procedure to overcome the difficulty in the analysis of unculturable species. The procedure has been tested on 6 Stichotrichia strains and uncultured Levicoleps biwae cells, targeting 18S rRNA gene sequences, after carrying out microscopic observations and obtaining photographic records. This procedure enables the systematic analysis of unculturable ciliate strains in natural environments by linking the morphology and genotype of a single cell.<br>

Journal

• Microbes and Environments

  Microbes and Environments 23 (4), 356-359, 2008

  Japanese Society of Microbial Ecology / Japanese Society of Soil Microbiology / Taiwan Society of Microbial Ecology / Japanese Society of Plant Microbe Interactions / Japanese Society for Extremophiles

Keywords

• Ciliates
• single-cell PCR
• 18S rRNA gene
• uncultured cell
• light microscopic analysis

Details

• CRID: 1390282679323030016
• NII Article ID: 110007005111
• NII Book ID: AA11551577
• DOI: 10.1264/jsme2.me08532
• ISSN: 13474405
• ISSN: 13426311
• NDL BIB ID: 9727649
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I9727649
• Web Site: http://www.jstage.jst.go.jp/article/jsme2/23/4/23_ME08532/_pdf
• Text Lang: en
• Data Source:
  • JaLC
  • NDL
  • Crossref
  • CiNii Articles
• Abstract License Flag: Disallowed