Real-time PCR method for detection of short DNA using a deoxyuridine probe and application for detection of fomivirsen
This study sought to develop a short DNA detection method using a deoxyuridine probe and polymerase chain reaction. The probe was hybridized to the target short DNA, which was then extended by DNA polymerase. The extended DNA was used for real-time PCR after the probe was removed by uracil DNA glyco...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Future Science Ltd
2022-12-01
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| Series: | BioTechniques |
| Subjects: | |
| Online Access: | https://www.future-science.com/doi/10.2144/btn-2022-0068 |
| _version_ | 1797980580852269056 |
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| author | Naoki Harikai Yuko Tanaka Satoshi Miyashita Kazumasa Zaima Kazufusa Shinomiya |
| author_facet | Naoki Harikai Yuko Tanaka Satoshi Miyashita Kazumasa Zaima Kazufusa Shinomiya |
| author_sort | Naoki Harikai |
| collection | DOAJ |
| description | This study sought to develop a short DNA detection method using a deoxyuridine probe and polymerase chain reaction. The probe was hybridized to the target short DNA, which was then extended by DNA polymerase. The extended DNA was used for real-time PCR after the probe was removed by uracil DNA glycosylase. This method measured from 0.01 to 10 nM of a model short DNA sequence of 17 nucleotides. The method was then used to detect the nucleic acid medicine fomivirsen, as well as 21 phosphorothioate nucleotides, and to quantify 0.1–100 nM of fomivirsen. This method may be useful for detecting short DNA fragments, such as functional nucleotides. |
| first_indexed | 2024-04-11T05:56:19Z |
| format | Article |
| id | doaj.art-5d89c9131e0d4518a724bc4c0a12bf1d |
| institution | Directory Open Access Journal |
| issn | 0736-6205 1940-9818 |
| language | English |
| last_indexed | 2024-04-11T05:56:19Z |
| publishDate | 2022-12-01 |
| publisher | Future Science Ltd |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj.art-5d89c9131e0d4518a724bc4c0a12bf1d2022-12-22T04:41:52ZengFuture Science LtdBioTechniques0736-62051940-98182022-12-0173628128710.2144/btn-2022-0068Real-time PCR method for detection of short DNA using a deoxyuridine probe and application for detection of fomivirsenNaoki Harikai0Yuko Tanaka1Satoshi Miyashita2Kazumasa Zaima3Kazufusa Shinomiya41School of Pharmacy, Nihon University, 7-7-1 Narashinodai, Funabashi, Chiba, 274-8555, Japan1School of Pharmacy, Nihon University, 7-7-1 Narashinodai, Funabashi, Chiba, 274-8555, Japan1School of Pharmacy, Nihon University, 7-7-1 Narashinodai, Funabashi, Chiba, 274-8555, Japan1School of Pharmacy, Nihon University, 7-7-1 Narashinodai, Funabashi, Chiba, 274-8555, Japan1School of Pharmacy, Nihon University, 7-7-1 Narashinodai, Funabashi, Chiba, 274-8555, JapanThis study sought to develop a short DNA detection method using a deoxyuridine probe and polymerase chain reaction. The probe was hybridized to the target short DNA, which was then extended by DNA polymerase. The extended DNA was used for real-time PCR after the probe was removed by uracil DNA glycosylase. This method measured from 0.01 to 10 nM of a model short DNA sequence of 17 nucleotides. The method was then used to detect the nucleic acid medicine fomivirsen, as well as 21 phosphorothioate nucleotides, and to quantify 0.1–100 nM of fomivirsen. This method may be useful for detecting short DNA fragments, such as functional nucleotides.https://www.future-science.com/doi/10.2144/btn-2022-0068fomivirsennucleic acid medicinepolymerase chain reactionshort DNAuracil DNA glycosylase |
| spellingShingle | Naoki Harikai Yuko Tanaka Satoshi Miyashita Kazumasa Zaima Kazufusa Shinomiya Real-time PCR method for detection of short DNA using a deoxyuridine probe and application for detection of fomivirsen BioTechniques fomivirsen nucleic acid medicine polymerase chain reaction short DNA uracil DNA glycosylase |
| title | Real-time PCR method for detection of short DNA using a deoxyuridine probe and application for detection of fomivirsen |
| title_full | Real-time PCR method for detection of short DNA using a deoxyuridine probe and application for detection of fomivirsen |
| title_fullStr | Real-time PCR method for detection of short DNA using a deoxyuridine probe and application for detection of fomivirsen |
| title_full_unstemmed | Real-time PCR method for detection of short DNA using a deoxyuridine probe and application for detection of fomivirsen |
| title_short | Real-time PCR method for detection of short DNA using a deoxyuridine probe and application for detection of fomivirsen |
| title_sort | real time pcr method for detection of short dna using a deoxyuridine probe and application for detection of fomivirsen |
| topic | fomivirsen nucleic acid medicine polymerase chain reaction short DNA uracil DNA glycosylase |
| url | https://www.future-science.com/doi/10.2144/btn-2022-0068 |
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