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Phylogenetic analysis of single bacterial cells contained in Antarctic iceberg.
Polar Science
Transdisciplinary Research Integration Center
Direct PCR amplification of the 16S rRNA gene from single microbial cells isolated from an Antarctic iceberg using laser microdissection microscopy
Katsuhiko Yanagihara, Hironori Niki, H. and Tomoya Baba
Polar Science (2011) Volume 5, Issue 3, September 2011, pp. 375-382
DOI: 10.1016/j.polar.2011.06.001

Here, we describe a technique that allows the genetic linage analysis of 16S rRNA genes in bacteria observed under a microscope. The technique includes the isolation of microbial cells using a laser microdissection microscope, lysis of the cells, and amplification of the 16S rRNA genes in the isolated cells without interference by bacterial DNA contamination from the experimental environment or reagents. Using this technique, we successfully determined 15 16S rRNA gene sequences in cells isolated from an Antarctic iceberg. These sequences showed 94%–100% identity to their closest strains, which included bacteria that occur in aqueous, marine, and soil environments.

A. Outline of the experiments. B, C. Microscopic images of the bacteria which were isolated and phylogenetically analyzed. The figures in parentheses show the 16SrRNA sequence identities to the known species.