【摘要】：重組酶聚合酶介導(dǎo)的等溫?cái)U(kuò)增(recombinase polymerase amplification,RPA)作為一種新型等溫?cái)U(kuò)增技術(shù),具有反應(yīng)時(shí)間短、靈敏度高、特異性強(qiáng)等特點(diǎn),在轉(zhuǎn)基因檢測(cè)領(lǐng)域得到了廣泛關(guān)注。本研究針對(duì)轉(zhuǎn)基因玉米Bt11外源插入鏈接區(qū)域設(shè)計(jì)篩選了特異性引物和探針,建立了快速準(zhǔn)確的熒光RPA檢測(cè)方法,在39℃條件下,20 min內(nèi)完成擴(kuò)增并且能夠?qū)崟r(shí)監(jiān)測(cè),可以準(zhǔn)確有效地檢測(cè)到50個(gè)拷貝的靶標(biāo)片段。而且,對(duì)于背景復(fù)雜的植物基因組DNA能夠得到準(zhǔn)確的擴(kuò)增曲線。本研究不僅可以滿足常規(guī)實(shí)驗(yàn)室檢測(cè),通過(guò)RPA便攜式儀器,還可以滿足轉(zhuǎn)基因植物現(xiàn)場(chǎng)檢測(cè),具有廣闊的應(yīng)用前景。
[Abstract]:As a new isothermal amplification technique, recombinant enzyme polymerase mediated isothermal amplification (recombinase polymerase amplification,RPA) has been widely concerned in the field of transgenic detection because of its short reaction time, high sensitivity and strong specificity. In this study, specific primers and probes were designed and screened for the exogenous inserted linked region of Bt11 in transgenic maize, and a rapid and accurate fluorescent RPA detection method was established. The PCR amplification was completed within 20 min at 39 鈩，

本文編號(hào)：2298979