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 應用微珠微流道結合動態等位基因特異性雜交法進行單核苷酸多態性檢測
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刊登日期：2014/05/21 |
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| ‧ 專利名稱 |
應用微珠微流道結合動態等位基因特異性雜交法進行單核苷酸多態性檢測 |
| ‧ 公開號 |
201410871
US20140080722
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| ‧ 專利權人 |
國立臺灣大學 |
‧ 專利國家
(申請日) |
中華民國 (2012/09/06)
美國 (2012/09/14)
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| ‧ 發明人/PI |
盧彥文,高培鈞,丁詩同,林恩仲,
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| ‧ 單位 |
生物產業機電工程學系
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| ‧ 簡歷/Experience |
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| 技術摘要 / Our Technology： |
本發明提供一種藉由利用動態等位基因特異性雜交技術與微珠微流道結合進行SNP檢測之方法，包含下列步驟：(a)將標的單股DNA固定至微珠上；(b)將標的單股DNA與等位基因特異性探針雜交；(c)將螢光標記插入標的探針雙鏈體(target-probe duplex)區域；(d)將微珠傳送至微流道；(e)將微珠加熱以使從步驟(c)所獲得的兩股的DNA分離(denature)；(f)監測於步驟(e)期間DNA的螢光強度以獲得熔解曲線；及(g)藉由熔解曲線分析法來測定單核苷酸多態性。同時，本發明提供具備試劑最小量的快速測定基因型之流程，其透過將微珠局限於所設計的流道性捕捉器內且透過溫度控制平台來控制進行熔解曲線分析。
The present invention provides a method of SNP detection by using DASH technique in bead-based microfluidics comprising following steps: (a) immobilizing a target single-strand DNA onto a microbead; (b) hybridizing the target single-strand DNA with an allele-specific probe; (c) intercalating a dye into a target-probe duplex region; (d) delivering the microbead into a microchannel; (e) heating the microbead to denature a hybridized DNA obtained from the step (c); (f) monitoring a fluorescence intensity of the hybridized DNA during the step (e) to obtain a melting curve; and (g) determining the SNP by a melting curve analysis method. Also, the present invention offers a rapid genotyping detection scheme with minimal amount of the reagents by confining the microbeads into designed fluidic traps and performing melting curve analysis controlled by a temperature control platform. The trapping mechanism was validated and optimized.
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| 專利簡述 / Intellectual Properties： |
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聯繫方式 / Contact： |
| 臺大產學合作總中心 / Center of Industry-Academia Collaboration, NTU |
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| Email：ordiac@ntu.edu.tw |
電話/Tel：02-3366-9945 |
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