本文選題：轉(zhuǎn)基因苜蓿KK- + 品系特異性　； 參考：《中國(guó)飼料》2017年14期

【摘要】：根據(jù)轉(zhuǎn)基因低木質(zhì)素苜蓿草品系KK179-2 5’端外源插入序列與苜蓿草基因組DNA之間的鄰接區(qū)序列設(shè)計(jì)引物和探針,建立了KK179-2品系特異性實(shí)時(shí)熒光PCR檢測(cè)方法,并對(duì)其特異性、靈敏度及可重復(fù)性進(jìn)行了測(cè)定。結(jié)果表明:建立的定量實(shí)時(shí)熒光PCR檢測(cè)方法特異性良好;標(biāo)準(zhǔn)曲線線性相關(guān)系數(shù)(R~2)為0.99,擴(kuò)增效率E為105%;檢測(cè)限為20拷貝,定量限為200拷貝。重復(fù)性實(shí)驗(yàn)顯示本方法的標(biāo)準(zhǔn)偏差(SD)及相對(duì)標(biāo)準(zhǔn)偏差(RSD)都在可接受范圍內(nèi)。綜上,建立的KK179-2品系特異性定量PCR檢測(cè)方法適于快速、準(zhǔn)確、穩(wěn)定地對(duì)轉(zhuǎn)基因苜蓿草KK179-2品系進(jìn)行檢測(cè)。
[Abstract]:The primers and probes were designed according to the sequence of the adjacent region between the exogenous insertion sequence at the end of KK179-2 5'terminal and the genomic DNA of alfalfa genome of transgenic alfalfa line KK179-2, and the specific real-time fluorescent PCR detection method for KK179-2 strain was established and its specificity was analyzed. Sensitivity and repeatability were determined. The results showed that the specificity of the method was good, the linear correlation coefficient of standard curve was 0.99, the amplification efficiency E was 105, the detection limit was 20 copies and the quantitative limit was 200 copies. Repeatability experiments showed that the standard deviation (SD) and relative standard deviation (RSD) of the method were within acceptable range. In conclusion, the established KK179-2 strain specific quantitative PCR method is suitable for rapid, accurate and stable detection of transgenic alfalfa line KK179-2.
【作者單位】： 黃埔出入境檢驗(yàn)檢疫局;廣東出入境檢驗(yàn)檢疫局;
【基金】：廣東出入境檢驗(yàn)檢疫局科技項(xiàng)目(2017GDK41、2016GDK60)
【分類號(hào)】：S541.9

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 劉彥泓;夏元鳳;陸旭先;楊滴;劉岑杰;張建寧;賀峰;;飼料中雞源性成分的實(shí)時(shí)熒光PCR檢測(cè)[J];飼料研究;2010年05期

2 周海亮;蔡麗;侯明生;;水稻細(xì)菌性基腐病病菌實(shí)時(shí)熒光PCR檢測(cè)[J];中國(guó)植保導(dǎo)刊;2013年11期

3 李樹(shù)清,易建平,李健,胡永強(qiáng),王巧全,陳志飛,周筱華,潘曉鐘,羅滿林,陳敏;胸膜肺炎放線桿菌的實(shí)時(shí)熒光PCR檢測(cè)[J];畜牧獸醫(yī)學(xué)報(bào);2005年09期

4 懷雁;徐麗慧;余山紅;謝關(guān)林;;水稻細(xì)菌性谷枯病菌的實(shí)時(shí)熒光PCR檢測(cè)技術(shù)研究[J];中國(guó)水稻科學(xué);2009年01期

5 杜雄偉;李葉;張雪;;傳染性支氣管炎病毒實(shí)時(shí)熒光PCR檢測(cè)方法的建立[J];食品科學(xué);2012年24期

6 張舒亞;韓麗;于翠;張坤;姚靜梅;張燕;;食品和飼料中貓?jiān)葱猿煞謱?shí)時(shí)熒光PCR檢測(cè)方法[J];上海農(nóng)業(yè)學(xué)報(bào);2013年04期

7 王，

本文編號(hào)：2054694