Bacteriophage strain typing by rapid single molecule analysis.
(2015) In Nucleic Acids Research 43(18). p.117-117- 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... (More)
- 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. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/5441960
- author
- Grunwald, Assaf ; Dahan, Moran ; Giesbertz, Anna ; Nilsson, Adam LU ; Nyberg, Lena K ; Weinhold, Elmar ; Ambjörnsson, Tobias LU ; Westerlund, Fredrik and Ebenstein, Yuval
- organization
- publishing date
- 2015
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Nucleic Acids Research
- volume
- 43
- issue
- 18
- pages
- 117 - 117
- publisher
- Oxford University Press
- external identifiers
-
- pmid:26019180
- wos:000366406500003
- scopus:84983802912
- pmid:26019180
- ISSN
- 1362-4962
- DOI
- 10.1093/nar/gkv563
- language
- English
- LU publication?
- yes
- id
- 4dc9c2a9-6488-4464-abd3-0aa02cdc253b (old id 5441960)
- date added to LUP
- 2016-04-01 09:51:18
- date last changed
- 2024-02-20 22:21:49
@article{4dc9c2a9-6488-4464-abd3-0aa02cdc253b, 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.}}, author = {{Grunwald, Assaf and Dahan, Moran and Giesbertz, Anna and Nilsson, Adam and Nyberg, Lena K and Weinhold, Elmar and Ambjörnsson, Tobias and Westerlund, Fredrik and Ebenstein, Yuval}}, issn = {{1362-4962}}, language = {{eng}}, number = {{18}}, pages = {{117--117}}, publisher = {{Oxford University Press}}, series = {{Nucleic Acids Research}}, title = {{Bacteriophage strain typing by rapid single molecule analysis.}}, url = {{http://dx.doi.org/10.1093/nar/gkv563}}, doi = {{10.1093/nar/gkv563}}, volume = {{43}}, year = {{2015}}, }