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Applied and Environmental Microbiology, April 2007, p. 2324-2328, Vol. 73, No. 7
0099-2240/07/$08.00+0     doi:10.1128/AEM.02038-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

SHORT REPORT

Detection of Low-Copy-Number Genomic DNA Sequences in Individual Bacterial Cells by Using Peptide Nucleic Acid-Assisted Rolling-Circle Amplification and Fluorescence In Situ Hybridization{triangledown} ,{dagger}

Irina Smolina,1 Charles Lee,2 and Maxim Frank-Kamenetskii1*

Center for Advanced Biotechnology and Department of Biomedical Engineering, Boston University, 36 Cummington St., Boston, Massachusetts 02215,1 Department of Pathology, Brigham and Women's Hospital, Harvard Medical School, 20 Shattuck St., Thorn Building, Boston, Massachusetts 021152

Received 28 August 2006/ Accepted 30 January 2007

An approach is proposed for in situ detection of short signature DNA sequences present in single copies per bacterial genome. The site is locally opened by peptide nucleic acids, and a circular oligonucleotide is assembled. The amplicon generated by rolling circle amplification is detected by hybridization with fluorescently labeled decorator probes.

* Corresponding author. Mailing address: Center for Advanced Biotechnology and Department of Biomedical Engineering, Boston University, 36 Cummington St., Boston, MA 02215. Phone: (617) 353-8492. Fax: (617) 353-8501. E-mail: mfk{at}bu.edu.

{triangledown} Published ahead of print on 9 February 2007.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, April 2007, p. 2324-2328, Vol. 73, No. 7
0099-2240/07/$08.00+0     doi:10.1128/AEM.02038-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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