Direct PCR amplification of the 16S rRNA gene from single microbial cells isolated from an Antarctic iceberg using laser microdissection microscopy

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.

► Microbial cells are isolated by using laser microdissection microscope. ► 16S rRNA genes are amplified from single microbial cell. ► Identification of bacteria included in Antarctic iceberg is demonstrated.