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Scalable amplification of strand subsets from chip-synthesized oligonucleotide libraries

Reference

Thorsten L. Schmidt, Brian J. Beliveau, Yavuz O. Uca, Mark Theilmann, Felipe Da Cruz, Chao-Ting Wu, William M. Shih, "Scalable amplification of strand subsets from chip-synthesized oligonucleotide libraries" , In Nat Commun, vol. 6, pp. 8634, Nov 2015. [doi]

Abstract

Synthetic oligonucleotides are the main cost factor for studies in DNA nanotechnology, genetics and synthetic biology, which all require thousands of these at high quality. Inexpensive chip-synthesized oligonucleotide libraries can contain hundreds of thousands of distinct sequences, however only at sub-femtomole quantities per strand. Here we present a selective oligonucleotide amplification method, based on three rounds of rolling-circle amplification, that produces nanomole amounts of single-stranded oligonucleotides per millilitre reaction. In a multistep one-pot procedure, subsets of hundreds or thousands of single-stranded DNAs with different lengths can selectively be amplified and purified together. These oligonucleotides are used to fold several DNA nanostructures and as primary fluorescence in situ hybridization probes. The amplification cost is lower than other reported methods (typically around US\$ 20 per nanomole total oligonucleotides produced) and is dominated by the use of commercial enzymes.

Bibtex

@article{schmidt_scalable_2015,
title={Scalable amplification of strand subsets from chip-synthesized oligonucleotide libraries},
volume={6},
copyright={© 2015 Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved.},
url={http://www.nature.com/ncomms/2015/151116/ncomms9634/full/ncomms9634.html},
doi={10.1038/ncomms9634},
abstract={Synthetic oligonucleotides are the main cost factor for studies in DNA nanotechnology, genetics and synthetic biology, which all require thousands of these at high quality. Inexpensive chip-synthesized oligonucleotide libraries can contain hundreds of thousands of distinct sequences, however only at sub-femtomole quantities per strand. Here we present a selective oligonucleotide amplification method, based on three rounds of rolling-circle amplification, that produces nanomole amounts of single-stranded oligonucleotides per millilitre reaction. In a multistep one-pot procedure, subsets of hundreds or thousands of single-stranded DNAs with different lengths can selectively be amplified and purified together. These oligonucleotides are used to fold several DNA nanostructures and as primary fluorescence in situ hybridization probes. The amplification cost is lower than other reported methods (typically around US\$ 20 per nanomole total oligonucleotides produced) and is dominated by the use of commercial enzymes.},
language={en},
urldate={2015-11-17},
journal={Nat Commun},
author={Schmidt, Thorsten L. and Beliveau, Brian J. and Uca, Yavuz O. and Theilmann, Mark and Da Cruz, Felipe and Wu, Chao-Ting and Shih, William M.},
month=nov,
year={2015},
note={00000},
keywords={Biological sciences, biotechnology, molecular biology, Nanotechnology},
pages={8634},
file={Schmidt et al. - 2015 - Scalable amplification of strand subsets from chip.pdf:C\:\\Users\\Thorsten\\Documents\\Zotero Bib\\storage\\M9SSUB4Z\\Schmidt et al. - 2015 - Scalable amplification of strand subsets from chip.pdf:application/pdf}
}

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