Sökning: WFRF:(Jeffet Jonathan)
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Super-Resolution Ge...
Super-Resolution Genome Mapping in Silicon Nanochannels
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- Jeffet, Jonathan (författare)
- Tel-Aviv University,Tel Aviv University
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- Kobo, Asaf (författare)
- Tel-Aviv University,Tel Aviv University
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- Su, Tianxiang (författare)
- Harvard University
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- Grunwald, Assaf (författare)
- Tel-Aviv University,Tel Aviv University
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- Green, Ori (författare)
- Tel-Aviv University,Tel Aviv University
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- Kinos, Adam (författare)
- Lund University,Lunds universitet,Atomfysik,Fysiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Atomic Physics,Department of Physics,Departments at LTH,Faculty of Engineering, LTH
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- Eisenberg, Eli (författare)
- Tel-Aviv University,Tel Aviv University,Rheinisch-Westfaelische Technische Hochschule Aachen,RWTH Aachen University
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- Ambjörnsson, Tobias (författare)
- Lund University,Lunds universitet,Beräkningsbiologi och biologisk fysik - Genomgår omorganisation,Institutionen för astronomi och teoretisk fysik - Genomgår omorganisation,Naturvetenskapliga fakulteten,Computational Biology and Biological Physics - Undergoing reorganization,Department of Astronomy and Theoretical Physics - Undergoing reorganization,Faculty of Science
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- Westerlund, Fredrik, 1978 (författare)
- Chalmers University of Technology,Chalmers tekniska högskola
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- Weinhold, Elmar (författare)
- RWTH Aachen University
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- Shabat, Doron (författare)
- Tel-Aviv University,Tel Aviv University
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- Purohit, Prashant K. (författare)
- University of Pennsylvania
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- Ebenstein, Yuval (författare)
- Tel-Aviv University,Tel Aviv University
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- Nilsson, A. (författare)
- Lunds universitet,Lund University
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(creator_code:org_t)
- 2016-09-23
- 2016
- Engelska 8 s.
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Ingår i: ACS Nano. - : American Chemical Society (ACS). - 1936-0851 .- 1936-086X. ; 10:11, s. 9823-9830
- Relaterad länk:
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http://dx.doi.org/10...
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https://lup.lub.lu.s...
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https://doi.org/10.1...
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https://research.cha...
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Abstract
Ämnesord
Stäng
- Optical genome mapping in nanochannels is a powerful genetic analysis method, complementary to deoxyribonucleic acid (DNA) sequencing. The method is based on detecting a pattern of fluorescent labels attached along individual DNA molecules. When such molecules are extended in nanochannels, the labels create a fluorescent genetic barcode that is used for mapping the DNA molecule to its genomic locus and identifying large-scale variation from the genome reference. Mapping resolution is currently limited by two main factors: the optical diffraction limit and the thermal fluctuations of DNA molecules suspended in the nanochannels. Here, we utilize single-molecule tracking and super-resolution localization in order to improve the mapping accuracy and resolving power of this genome mapping technique and achieve a 15-fold increase in resolving power compared to currently practiced methods. We took advantage of a naturally occurring genetic repeat array and labeled each repeat with custom-designed Trolox conjugated fluorophores for enhanced photostability. This model system allowed us to acquire extremely long image sequences of the equally spaced fluorescent markers along DNA molecules, enabling detailed characterization of nanoconfined DNA dynamics and quantitative comparison to the Odijk theory for confined polymer chains. We present a simple method to overcome the thermal fluctuations in the nanochannels and exploit single-step photobleaching to resolve subdiffraction spaced fluorescent markers along fluctuating DNA molecules with ∼100 bp resolution. In addition, we show how time-averaging over just ∼50 frames of 40 ms enhances mapping accuracy, improves mapping P-value scores by 3 orders of magnitude compared to nonaveraged alignment, and provides a significant advantage for analyzing structural variations between DNA molecules with similar sequence composition.
Ämnesord
- TEKNIK OCH TEKNOLOGIER -- Nanoteknik -- Nanoteknik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Nano-technology -- Nano-technology (hsv//eng)
- NATURVETENSKAP -- Biologi -- Genetik (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Genetics (hsv//eng)
- NATURVETENSKAP -- Biologi -- Biofysik (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biophysics (hsv//eng)
- NATURVETENSKAP -- Fysik -- Annan fysik (hsv//swe)
- NATURAL SCIENCES -- Physical Sciences -- Other Physics Topics (hsv//eng)
- TEKNIK OCH TEKNOLOGIER -- Nanoteknik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Nano-technology (hsv//eng)
Nyckelord
- confined polymers
- DNA labeling
- nanochannels
- optical genome mapping
- single-molecule
- super-resolution
- super-resolution
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ACS Nano
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Jeffet, Jonathan
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Kobo, Asaf
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Su, Tianxiang
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Grunwald, Assaf
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Green, Ori
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Kinos, Adam
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visa fler...
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Eisenberg, Eli
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Ambjörnsson, Tob ...
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Westerlund, Fred ...
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Weinhold, Elmar
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Shabat, Doron
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Purohit, Prashan ...
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Ebenstein, Yuval
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Nilsson, A.
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visa färre...
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- TEKNIK OCH TEKNOLOGIER
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TEKNIK OCH TEKNO ...
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och Nanoteknik
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och Nanoteknik
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NATURVETENSKAP
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NATURVETENSKAP
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och Biologi
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och Biofysik
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- NATURVETENSKAP
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NATURVETENSKAP
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och Annan fysik
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ACS Nano
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