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Microarray Generation of Thousand-Member Oligonucleotide Libraries

The ability to efficiently and economically generate libraries of defined pieces of DNA would have a myriad of applications, not least in the area of defined or directed sequencing and synthetic biology, but also in applications associated with encoding and tagging. In this manuscript DNA microarray...

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Autores principales: Svensen, Nina, Díaz-Mochón, Juan José, Bradley, Mark
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3179494/
https://www.ncbi.nlm.nih.gov/pubmed/21966380
http://dx.doi.org/10.1371/journal.pone.0024906
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author Svensen, Nina
Díaz-Mochón, Juan José
Bradley, Mark
author_facet Svensen, Nina
Díaz-Mochón, Juan José
Bradley, Mark
author_sort Svensen, Nina
collection PubMed
description The ability to efficiently and economically generate libraries of defined pieces of DNA would have a myriad of applications, not least in the area of defined or directed sequencing and synthetic biology, but also in applications associated with encoding and tagging. In this manuscript DNA microarrays were used to allow the linear amplification of immobilized DNA sequences from the array followed by PCR amplification. Arrays of increasing sophistication (1, 10, 3,875, 10,000 defined sequences) were used to validate the process, with sequences verified by selective hybridization to a complementary DNA microarray and DNA sequencing, which demonstrated a PCR error rate of 9.7×10(−3)/site/duplication. This technique offers an economical and efficient way of producing specific DNA libraries of hundreds to thousands of members with the DNA-arrays being used as “factories” allowing specific DNA oligonucleotide pools to be generated. We also found substantial variance observed between the sequence frequencies found via Solexa sequencing and microarray analysis, highlighting the care needed in the interpretation of profiling data.
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spelling pubmed-31794942011-09-30 Microarray Generation of Thousand-Member Oligonucleotide Libraries Svensen, Nina Díaz-Mochón, Juan José Bradley, Mark PLoS One Research Article The ability to efficiently and economically generate libraries of defined pieces of DNA would have a myriad of applications, not least in the area of defined or directed sequencing and synthetic biology, but also in applications associated with encoding and tagging. In this manuscript DNA microarrays were used to allow the linear amplification of immobilized DNA sequences from the array followed by PCR amplification. Arrays of increasing sophistication (1, 10, 3,875, 10,000 defined sequences) were used to validate the process, with sequences verified by selective hybridization to a complementary DNA microarray and DNA sequencing, which demonstrated a PCR error rate of 9.7×10(−3)/site/duplication. This technique offers an economical and efficient way of producing specific DNA libraries of hundreds to thousands of members with the DNA-arrays being used as “factories” allowing specific DNA oligonucleotide pools to be generated. We also found substantial variance observed between the sequence frequencies found via Solexa sequencing and microarray analysis, highlighting the care needed in the interpretation of profiling data. Public Library of Science 2011-09-23 /pmc/articles/PMC3179494/ /pubmed/21966380 http://dx.doi.org/10.1371/journal.pone.0024906 Text en Svensen et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Svensen, Nina
Díaz-Mochón, Juan José
Bradley, Mark
Microarray Generation of Thousand-Member Oligonucleotide Libraries
title Microarray Generation of Thousand-Member Oligonucleotide Libraries
title_full Microarray Generation of Thousand-Member Oligonucleotide Libraries
title_fullStr Microarray Generation of Thousand-Member Oligonucleotide Libraries
title_full_unstemmed Microarray Generation of Thousand-Member Oligonucleotide Libraries
title_short Microarray Generation of Thousand-Member Oligonucleotide Libraries
title_sort microarray generation of thousand-member oligonucleotide libraries
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3179494/
https://www.ncbi.nlm.nih.gov/pubmed/21966380
http://dx.doi.org/10.1371/journal.pone.0024906
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