Bacteriophage strain typing by rapid single molecule analysis.

Abstract:

:Rapid characterization of unknown biological samples is under the focus of many current studies. Here we report a method for screening of biological samples by optical mapping of their DNA. We use a novel, one-step chemo-enzymatic reaction to covalently bind fluorophores to DNA at the four-base recognition sites of a DNA methyltransferase. Due to the diffraction limit of light, the dense distribution of labels results in a continuous fluorescent signal along the DNA. The amplitude modulations (AM) of the fluorescence intensity along the stretched DNA molecules exhibit a unique molecular fingerprint that can be used for identification. We show that this labelling scheme is highly informative, allowing accurate genotyping. We demonstrate the method by labelling the genomes of λ and T7 bacteriophages, resulting in a consistent, unique AM profile for each genome. These profiles are also successfully used for identification of the phages from a background phage library. Our method may provide a facile route for screening and typing of various organisms and has potential applications in metagenomics studies of various ecosystems.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Grunwald A,Dahan M,Giesbertz A,Nilsson A,Nyberg LK,Weinhold E,Ambjörnsson T,Westerlund F,Ebenstein Y

doi

10.1093/nar/gkv563

subject

Has Abstract

pub_date

2015-10-15 00:00:00

pages

e117

issue

18

eissn

0305-1048

issn

1362-4962

pii

gkv563

journal_volume

43

pub_type

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