A Novel Helper qPCR Assay for the Detection of miRNA Using Target/Helper Template for Primer Formation
A novel, simple, and sensitive quantitative polymerase chain reaction (qPCR) technology, which is termed as helper qPCR, was established to detect miRNA. In this assay, the target miRNA sequence was introduced as helper template for a reaction switch preforming two-step real-time qPCR strategy. Firs...
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| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wiley
2022-01-01
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| Series: | International Journal of Analytical Chemistry |
| Online Access: | http://dx.doi.org/10.1155/2022/6918054 |
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| _version_ | 1850174041624674304 |
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| author | Xianfeng Jiang Jiahui Wang Xudan Shen Jadera Talap Yang Yang Dan Yang Guizhou Xiao Sheng Cai |
| author_facet | Xianfeng Jiang Jiahui Wang Xudan Shen Jadera Talap Yang Yang Dan Yang Guizhou Xiao Sheng Cai |
| author_sort | Xianfeng Jiang |
| collection | DOAJ |
| description | A novel, simple, and sensitive quantitative polymerase chain reaction (qPCR) technology, which is termed as helper qPCR, was established to detect miRNA. In this assay, the target miRNA sequence was introduced as helper template for a reaction switch preforming two-step real-time qPCR strategy. Firstly, the reverse primer was reverse transcribed to form “mediator primer” after binding to the target miRNA. Then, the mediator primer was further extended to form “active template” with annealing to the mediator template. In the end, the active template was amplified and detected by the qPCR reaction system with the help of reverse and forward primers. The SYBR Green dye was used for fluorescence quantification, which is quicker and cheaper than the fluorescent probes, as the detection limit of this assay was 1 pM. This helper qPCR system can be used for different miRNAs detection by redesigning reverse primer for target, indicating this strategy could afford good performance in detecting multiple miRNAs and has a promising application prospect. |
| format | Article |
| id | doaj-art-c7262c6ddc2b4714902a15e2c8005f34 |
| institution | OA Journals |
| issn | 1687-8779 |
| language | English |
| publishDate | 2022-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | International Journal of Analytical Chemistry |
| spelling | doaj-art-c7262c6ddc2b4714902a15e2c8005f342025-08-20T02:19:44ZengWileyInternational Journal of Analytical Chemistry1687-87792022-01-01202210.1155/2022/6918054A Novel Helper qPCR Assay for the Detection of miRNA Using Target/Helper Template for Primer FormationXianfeng Jiang0Jiahui Wang1Xudan Shen2Jadera Talap3Yang Yang4Dan Yang5Guizhou Xiao6Sheng Cai7Sir Run Run Shaw HospitalSir Run Run Shaw HospitalZhejiang Province Key Laboratory of Anti-Cancer Drug ResearchZhejiang Province Key Laboratory of Anti-Cancer Drug ResearchSir Run Run Shaw HospitalZhejiang Province Key Laboratory of Anti-Cancer Drug ResearchSir Run Run Shaw HospitalSir Run Run Shaw HospitalA novel, simple, and sensitive quantitative polymerase chain reaction (qPCR) technology, which is termed as helper qPCR, was established to detect miRNA. In this assay, the target miRNA sequence was introduced as helper template for a reaction switch preforming two-step real-time qPCR strategy. Firstly, the reverse primer was reverse transcribed to form “mediator primer” after binding to the target miRNA. Then, the mediator primer was further extended to form “active template” with annealing to the mediator template. In the end, the active template was amplified and detected by the qPCR reaction system with the help of reverse and forward primers. The SYBR Green dye was used for fluorescence quantification, which is quicker and cheaper than the fluorescent probes, as the detection limit of this assay was 1 pM. This helper qPCR system can be used for different miRNAs detection by redesigning reverse primer for target, indicating this strategy could afford good performance in detecting multiple miRNAs and has a promising application prospect.http://dx.doi.org/10.1155/2022/6918054 |
| spellingShingle | Xianfeng Jiang Jiahui Wang Xudan Shen Jadera Talap Yang Yang Dan Yang Guizhou Xiao Sheng Cai A Novel Helper qPCR Assay for the Detection of miRNA Using Target/Helper Template for Primer Formation International Journal of Analytical Chemistry |
| title | A Novel Helper qPCR Assay for the Detection of miRNA Using Target/Helper Template for Primer Formation |
| title_full | A Novel Helper qPCR Assay for the Detection of miRNA Using Target/Helper Template for Primer Formation |
| title_fullStr | A Novel Helper qPCR Assay for the Detection of miRNA Using Target/Helper Template for Primer Formation |
| title_full_unstemmed | A Novel Helper qPCR Assay for the Detection of miRNA Using Target/Helper Template for Primer Formation |
| title_short | A Novel Helper qPCR Assay for the Detection of miRNA Using Target/Helper Template for Primer Formation |
| title_sort | novel helper qpcr assay for the detection of mirna using target helper template for primer formation |
| url | http://dx.doi.org/10.1155/2022/6918054 |
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