Bacteriophage strain typing by rapid single molecule analysis.

Summary, in English

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.

• Author: Assaf Grunwald; Moran Dahan; Anna Giesbertz; Adam Nilsson; Lena K Nyberg; Elmar Weinhold; Tobias Ambjörnsson; Fredrik Westerlund; Yuval Ebenstein
• Department/s: Atomic Physics; Computational Biology and Biological Physics - Has been reorganised
• Publishing year: 2015
• Language: English
• Pages: 117-117
• Publication/Series: Nucleic Acids Research
• Volume: 43
• Issue: 18
• Links: http://dx.doi.org/10.1093/nar/gkv563
• Document type: Journal article
• Publisher: Oxford University Press
• Topic: Biophysics; Other Physics Topics
• Status: Published
• ISBN/ISSN/Other: ISSN: 1362-4962