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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Public Library of Science
2011
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| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3179494/ |
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pubmed-31794942011-09-30 Microarray Generation of Thousand-Member Oligonucleotide Libraries Svensen, Nina Díaz-Mochón, Juan José Bradley, Mark 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. |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
Svensen, Nina Díaz-Mochón, Juan José Bradley, Mark |
| spellingShingle |
Svensen, Nina Díaz-Mochón, Juan José Bradley, Mark Microarray Generation of Thousand-Member Oligonucleotide Libraries |
| author_facet |
Svensen, Nina Díaz-Mochón, Juan José Bradley, Mark |
| author_sort |
Svensen, Nina |
| title |
Microarray Generation of Thousand-Member Oligonucleotide Libraries |
| title_short |
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_sort |
microarray generation of thousand-member oligonucleotide libraries |
| 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. |
| publisher |
Public Library of Science |
| publishDate |
2011 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3179494/ |
| _version_ |
1611477183930302464 |