Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 546465
Title Linearization and Labeling of Single-Stranded DNA for Optical Sequence Analysis
Author(s) Basak, Rajib; Liu, Fan; Qureshi, Sarfraz; Gupta, Neelima; Zhang, Ce; Vries, Renko de; Kan, Jeroen A. van; Dheen, S.T.; Maarel, Johan R.C. van der
Source Journal of Physical Chemistry Letters 10 (2019)3. - ISSN 1948-7185 - p. 316 - 321.
DOI https://doi.org/10.1021/acs.jpclett.8b03465
Department(s) VLAG
Physical Chemistry and Soft Matter
Publication type Refereed Article in a scientific journal
Publication year 2019
Abstract

Genetic profiling would benefit from linearization of ssDNA through the exposure of the unpaired bases to gene-targeting probes. This is compromised by ssDNA's high flexibility and tendency to form self-annealed structures. Here, we demonstrate that self-annealing can be avoided through controlled coating with a cationic-neutral diblock polypeptide copolymer. Coating does not preclude site-specific binding of fluorescence labeled oligonucleotides. Bottlebrush-coated ssDNA can be linearized by confinement inside a nanochannel or molecular combing. A stretch of 0.32 nm per nucleotide is achieved inside a channel with a cross-section of 100 nm and a 2-fold excess of polypeptide with respect to DNA charge. With combing, the complexes are stretched to a similar extent. Atomic force microscopy of dried complexes on silica revealed that the contour and persistence lengths are close to those of dsDNA in the B-form. Labeling is based on hybridization and not limited by restriction enzymes. Enzyme-free labeling offers new opportunities for the detection of specific sequences.

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