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Label-Free and Sensitive Fluorescent Detection of Sequence-Specific Single-Strand DNA Based on S1 Nuclease Cleavage Effects
The ability to detect sequence-specific single-strand DNA (ssDNA) in complex, contaminant-ridden samples, using a fluorescent method directly without a DNA extraction and PCR step could simplify the detection of pathogens in the field and in the clinic. Here, we have demonstrated a simple label-free...
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/PMC4186753/ https://www.ncbi.nlm.nih.gov/pubmed/25285445 http://dx.doi.org/10.1371/journal.pone.0108401 |
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author | Guan, Zheng Liu, Jinchuan Bai, Wenhui Lv, Zhenzhen Jiang, Xiaoling Yang, Shuming Chen, Ailiang Lv, Guiyuan |
author_facet | Guan, Zheng Liu, Jinchuan Bai, Wenhui Lv, Zhenzhen Jiang, Xiaoling Yang, Shuming Chen, Ailiang Lv, Guiyuan |
author_sort | Guan, Zheng |
collection | PubMed |
description | The ability to detect sequence-specific single-strand DNA (ssDNA) in complex, contaminant-ridden samples, using a fluorescent method directly without a DNA extraction and PCR step could simplify the detection of pathogens in the field and in the clinic. Here, we have demonstrated a simple label-free sensing strategy to detect ssDNA by employing its complementary ssDNA, S1 nuclease and nucleic acid fluorescent dyes. Upon clearing away redundant complementary ssDNA and possibly mismatched double strand DNA by using S1 nuclease, the fluorescent signal-to-noise ratio could be increased dramatically. It enabled the method to be adaptable to three different types of DNA fluorescent dyes and the ability to detect target ssDNA in complex, multicomponent samples, like tissue homogenate. The method can distinguish a two-base mismatch from avian influenza A (H1N1) virus. Also, it can detect the appearance of 50 pM target ssDNA in 0.5 µg·mL(−1) Lambda DNA, and 50 nM target ssDNA in 5 µg·mL(−1) Lambda DNA or in tissue homogenate. It is facile and cost-effective, and could be easily extended to detect other ssDNA with many common nucleic acid fluorescent dyes. |
format | Online Article Text |
id | pubmed-4186753 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-41867532014-10-16 Label-Free and Sensitive Fluorescent Detection of Sequence-Specific Single-Strand DNA Based on S1 Nuclease Cleavage Effects Guan, Zheng Liu, Jinchuan Bai, Wenhui Lv, Zhenzhen Jiang, Xiaoling Yang, Shuming Chen, Ailiang Lv, Guiyuan PLoS One Research Article The ability to detect sequence-specific single-strand DNA (ssDNA) in complex, contaminant-ridden samples, using a fluorescent method directly without a DNA extraction and PCR step could simplify the detection of pathogens in the field and in the clinic. Here, we have demonstrated a simple label-free sensing strategy to detect ssDNA by employing its complementary ssDNA, S1 nuclease and nucleic acid fluorescent dyes. Upon clearing away redundant complementary ssDNA and possibly mismatched double strand DNA by using S1 nuclease, the fluorescent signal-to-noise ratio could be increased dramatically. It enabled the method to be adaptable to three different types of DNA fluorescent dyes and the ability to detect target ssDNA in complex, multicomponent samples, like tissue homogenate. The method can distinguish a two-base mismatch from avian influenza A (H1N1) virus. Also, it can detect the appearance of 50 pM target ssDNA in 0.5 µg·mL(−1) Lambda DNA, and 50 nM target ssDNA in 5 µg·mL(−1) Lambda DNA or in tissue homogenate. It is facile and cost-effective, and could be easily extended to detect other ssDNA with many common nucleic acid fluorescent dyes. Public Library of Science 2014-10-06 /pmc/articles/PMC4186753/ /pubmed/25285445 http://dx.doi.org/10.1371/journal.pone.0108401 Text en © 2014 Guan 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 Guan, Zheng Liu, Jinchuan Bai, Wenhui Lv, Zhenzhen Jiang, Xiaoling Yang, Shuming Chen, Ailiang Lv, Guiyuan Label-Free and Sensitive Fluorescent Detection of Sequence-Specific Single-Strand DNA Based on S1 Nuclease Cleavage Effects |
title | Label-Free and Sensitive Fluorescent Detection of Sequence-Specific Single-Strand DNA Based on S1 Nuclease Cleavage Effects |
title_full | Label-Free and Sensitive Fluorescent Detection of Sequence-Specific Single-Strand DNA Based on S1 Nuclease Cleavage Effects |
title_fullStr | Label-Free and Sensitive Fluorescent Detection of Sequence-Specific Single-Strand DNA Based on S1 Nuclease Cleavage Effects |
title_full_unstemmed | Label-Free and Sensitive Fluorescent Detection of Sequence-Specific Single-Strand DNA Based on S1 Nuclease Cleavage Effects |
title_short | Label-Free and Sensitive Fluorescent Detection of Sequence-Specific Single-Strand DNA Based on S1 Nuclease Cleavage Effects |
title_sort | label-free and sensitive fluorescent detection of sequence-specific single-strand dna based on s1 nuclease cleavage effects |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4186753/ https://www.ncbi.nlm.nih.gov/pubmed/25285445 http://dx.doi.org/10.1371/journal.pone.0108401 |
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