Accession Number : AD1042520


Title :   Scalable Amplification of Strand Subsets from Chip-Synthesized Oligonucleotide Libraries (Open Access)


Descriptive Note : Journal Article - Open Access


Corporate Author : DANA-FARBER CANCER INST BOSTON MA BOSTON United States


Personal Author(s) : Schmidt,Thorsten L ; Beliveau,Brian J ; Uca,Yavuz O ; Theilmann,Mark ; Da Cruz,Felipe ; Wu,Chao-Ting ; Shih,William M


Full Text : https://apps.dtic.mil/dtic/tr/fulltext/u2/1042520.pdf


Report Date : 16 Nov 2015


Pagination or Media Count : 8


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.


Descriptors :   DNA MICROARRAYS , nanotechnology , confocal laser scanning microscopy , gel electrophoresis , synthetic biology , Molecular biology , biotechnology , nucleotides , genetics


Subject Categories : Biomedical Instrumentation and Bioengineering


Distribution Statement : APPROVED FOR PUBLIC RELEASE