Bacteriophage strain typing by rapid single molecule analysis. Issue 18 (10th October 2015)

• Record Type: Journal Article
• Authors: Grunwald, Assaf; Dahan, Moran; Giesbertz, Anna; Nilsson, Adam; Nyberg, Lena K.; Weinhold, Elmar; Ambjörnsson, Tobias; Westerlund, Fredrik; Ebenstein, Yuval
• Is Part Of: Nucleic acids research. Volume 43:Issue 18(2015)
• Page Start: e117
• Page End: e117
• Publication Date: 2015-10-10
• Subjects: Nucleic acids -- Periodicals; Molecular biology -- Periodicals; 572.805
• DOI: 10.1093/nar/gkv563
• Languages: English
• ISSNs: 0305-1048

Abstract

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.