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By-Product Formation in Repetitive PCR Amplification of DNA Libraries during SELEX
The selection of nucleic acid aptamers is an increasingly important approach to generate specific ligands binding to virtually any molecule of choice. However, selection-inherent amplification procedures are prone to artificial by-product formation that prohibits the enrichment of target-recognizing...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4260880/ https://www.ncbi.nlm.nih.gov/pubmed/25490402 http://dx.doi.org/10.1371/journal.pone.0114693 |
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author | Tolle, Fabian Wilke, Julian Wengel, Jesper Mayer, Günter |
author_facet | Tolle, Fabian Wilke, Julian Wengel, Jesper Mayer, Günter |
author_sort | Tolle, Fabian |
collection | PubMed |
description | The selection of nucleic acid aptamers is an increasingly important approach to generate specific ligands binding to virtually any molecule of choice. However, selection-inherent amplification procedures are prone to artificial by-product formation that prohibits the enrichment of target-recognizing aptamers. Little is known about the formation of such by-products when employing nucleic acid libraries as templates. We report on the formation of two different forms of by-products, named ladder- and non-ladder-type observed during repetitive amplification in the course of in vitro selection experiments. Based on sequence information and the amplification behaviour of defined enriched nucleic acid molecules we suppose a molecular mechanism through which these amplification by-products are built. Better understanding of these mechanisms might help to find solutions minimizing by-product formation and improving the success rate of aptamer selection. |
format | Online Article Text |
id | pubmed-4260880 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-42608802014-12-15 By-Product Formation in Repetitive PCR Amplification of DNA Libraries during SELEX Tolle, Fabian Wilke, Julian Wengel, Jesper Mayer, Günter PLoS One Research Article The selection of nucleic acid aptamers is an increasingly important approach to generate specific ligands binding to virtually any molecule of choice. However, selection-inherent amplification procedures are prone to artificial by-product formation that prohibits the enrichment of target-recognizing aptamers. Little is known about the formation of such by-products when employing nucleic acid libraries as templates. We report on the formation of two different forms of by-products, named ladder- and non-ladder-type observed during repetitive amplification in the course of in vitro selection experiments. Based on sequence information and the amplification behaviour of defined enriched nucleic acid molecules we suppose a molecular mechanism through which these amplification by-products are built. Better understanding of these mechanisms might help to find solutions minimizing by-product formation and improving the success rate of aptamer selection. Public Library of Science 2014-12-09 /pmc/articles/PMC4260880/ /pubmed/25490402 http://dx.doi.org/10.1371/journal.pone.0114693 Text en © 2014 Tolle 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 Tolle, Fabian Wilke, Julian Wengel, Jesper Mayer, Günter By-Product Formation in Repetitive PCR Amplification of DNA Libraries during SELEX |
title | By-Product Formation in Repetitive PCR Amplification of DNA Libraries during SELEX |
title_full | By-Product Formation in Repetitive PCR Amplification of DNA Libraries during SELEX |
title_fullStr | By-Product Formation in Repetitive PCR Amplification of DNA Libraries during SELEX |
title_full_unstemmed | By-Product Formation in Repetitive PCR Amplification of DNA Libraries during SELEX |
title_short | By-Product Formation in Repetitive PCR Amplification of DNA Libraries during SELEX |
title_sort | by-product formation in repetitive pcr amplification of dna libraries during selex |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4260880/ https://www.ncbi.nlm.nih.gov/pubmed/25490402 http://dx.doi.org/10.1371/journal.pone.0114693 |
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