Bacteriophage strain typing by rapid single molecule analysis

Assaf Grunwald, Moran Dahan, Anna Giesbertz, Adam Nilsson, Lena K. Nyberg, Elmar Weinhold, Tobias Ambjörnsson, Fredrik Westerlund, Yuval Ebenstein

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Article numbere117
JournalNucleic Acids Research
Volume43
Issue number18
DOIs
StatePublished - 15 Oct 2015

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