Clone libraries and single cell genome amplification reveal extended diversity of uncultivated magnetotactic bacteria from marine and freshwater environments
Article first published online: 26 OCT 2012
© 2012 Society for Applied Microbiology and Blackwell Publishing Ltd
Special Issue: Marine Microbial Ecophysiology and Metagenomics
Volume 15, Issue 5, pages 1290–1301, May 2013
How to Cite
Kolinko, S., Wanner, G., Katzmann, E., Kiemer, F., M. Fuchs, B. and Schüler, D. (2013), Clone libraries and single cell genome amplification reveal extended diversity of uncultivated magnetotactic bacteria from marine and freshwater environments. Environmental Microbiology, 15: 1290–1301. doi: 10.1111/1462-2920.12004
- Issue published online: 18 APR 2013
- Article first published online: 26 OCT 2012
- Accepted manuscript online: 28 SEP 2012 06:01AM EST
- Manuscript Accepted: 21 SEP 2012
- Manuscript Revised: 7 SEP 2012
- Manuscript Received: 25 JUL 2012
- Deutsche Forschungsgemeinschaft. Grant Numbers: DFG Schu1080/9-1, 11-1
Fig. S1. Schematic of experimental set-up of the combined approach to recover diversity from magnetic collections and genomic analysis of single cells. After a magnetic enrichment from an environmental sample (A) a selective enriched collection of MTB was obtained (B). Using universal eubacterial primers almost full-length 16S rRNA genes were amplified and subcloned for sequencing (C). Every MTB morphotype was selected by single cell sorting applying micromanipulation under microscopic control (D). Genomic DNA of selected cells were amplified using the viral Φ29 DNA-polymerase by multiple displacement amplification (MDA). Amplified DNA served as template for 16S rRNA PCR allowing phylogenetic analysis of selected morphotypes (E). For genetic analysis of microsorted CS-05 cells genomic DNA was amplified a second time by the MDA, enzymatically debranched, checked for contaminations and subcloned for analysis (F).
Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.