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The Fidelity Index provides a systematic quantitation of star activity of DNA restriction endonucleases
Restriction endonucleases are the basic tools of molecular biology. Many restriction endonucleases show relaxed sequence recognition, called star activity, as an inherent property under various digestion conditions including the optimal ones. To quantify this property we propose the concept of the F...
Autores principales: | , , , , |
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Formato: | Texto |
Lenguaje: | English |
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Oxford University Press
2008
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2396408/ https://www.ncbi.nlm.nih.gov/pubmed/18413342 http://dx.doi.org/10.1093/nar/gkn182 |
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author | Wei, Hua Therrien, Caitlin Blanchard, Aine Guan, Shengxi Zhu, Zhenyu |
author_facet | Wei, Hua Therrien, Caitlin Blanchard, Aine Guan, Shengxi Zhu, Zhenyu |
author_sort | Wei, Hua |
collection | PubMed |
description | Restriction endonucleases are the basic tools of molecular biology. Many restriction endonucleases show relaxed sequence recognition, called star activity, as an inherent property under various digestion conditions including the optimal ones. To quantify this property we propose the concept of the Fidelity Index (FI), which is defined as the ratio of the maximum enzyme amount showing no star activity to the minimum amount needed for complete digestion at the cognate recognition site for any particular restriction endonuclease. Fidelity indices for a large number of restriction endonucleases are reported here. The effects of reaction vessel, reaction volume, incubation mode, substrate differences, reaction time, reaction temperature and additional glycerol, DMSO, ethanol and Mn(2+) on the FI are also investigated. The FI provides a practical guideline for the use of restriction endonucleases and defines a fundamental property by which restriction endonucleases can be characterized. |
format | Text |
id | pubmed-2396408 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-23964082008-05-28 The Fidelity Index provides a systematic quantitation of star activity of DNA restriction endonucleases Wei, Hua Therrien, Caitlin Blanchard, Aine Guan, Shengxi Zhu, Zhenyu Nucleic Acids Res Methods Online Restriction endonucleases are the basic tools of molecular biology. Many restriction endonucleases show relaxed sequence recognition, called star activity, as an inherent property under various digestion conditions including the optimal ones. To quantify this property we propose the concept of the Fidelity Index (FI), which is defined as the ratio of the maximum enzyme amount showing no star activity to the minimum amount needed for complete digestion at the cognate recognition site for any particular restriction endonuclease. Fidelity indices for a large number of restriction endonucleases are reported here. The effects of reaction vessel, reaction volume, incubation mode, substrate differences, reaction time, reaction temperature and additional glycerol, DMSO, ethanol and Mn(2+) on the FI are also investigated. The FI provides a practical guideline for the use of restriction endonucleases and defines a fundamental property by which restriction endonucleases can be characterized. Oxford University Press 2008-05 2008-04-15 /pmc/articles/PMC2396408/ /pubmed/18413342 http://dx.doi.org/10.1093/nar/gkn182 Text en © 2008 The Author(s) http://creativecommons.org/licenses/by-nc/2.0/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Methods Online Wei, Hua Therrien, Caitlin Blanchard, Aine Guan, Shengxi Zhu, Zhenyu The Fidelity Index provides a systematic quantitation of star activity of DNA restriction endonucleases |
title | The Fidelity Index provides a systematic quantitation of star activity of DNA restriction endonucleases |
title_full | The Fidelity Index provides a systematic quantitation of star activity of DNA restriction endonucleases |
title_fullStr | The Fidelity Index provides a systematic quantitation of star activity of DNA restriction endonucleases |
title_full_unstemmed | The Fidelity Index provides a systematic quantitation of star activity of DNA restriction endonucleases |
title_short | The Fidelity Index provides a systematic quantitation of star activity of DNA restriction endonucleases |
title_sort | fidelity index provides a systematic quantitation of star activity of dna restriction endonucleases |
topic | Methods Online |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2396408/ https://www.ncbi.nlm.nih.gov/pubmed/18413342 http://dx.doi.org/10.1093/nar/gkn182 |
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