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.
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.
Smolina
Irina
I
Center for Advanced Biotechnology and Department of Biomedical Engineering, Boston University, 36 Cummington St., Boston, MA 02215, USA.
Lee
Charles
C
Frank-Kamenetskii
Maxim
M
D001412:Bacillus subtilis / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000032:analysis* / Q000737:chemistry; D004926:Escherichia coli / Q000302:isolation & purification*; D018628:Gene Dosage; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D020135:Peptide Nucleic Acids / Q000235:genetics*; D017422:Sequence Analysis, DNA
D004269:DNA, Bacterial; D020135:Peptide Nucleic Acids