本文選題：轉(zhuǎn)基因苜蓿KK- + 品系特異性　； 參考：《中國飼料》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)計引物和探針,建立了KK179-2品系特異性實時熒光PCR檢測方法,并對其特異性、靈敏度及可重復(fù)性進行了測定。結(jié)果表明:建立的定量實時熒光PCR檢測方法特異性良好;標(biāo)準(zhǔn)曲線線性相關(guān)系數(shù)(R~2)為0.99,擴增效率E為105%;檢測限為20拷貝,定量限為200拷貝。重復(fù)性實驗顯示本方法的標(biāo)準(zhǔn)偏差(SD)及相對標(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檢測方法適于快速、準(zhǔn)確、穩(wěn)定地對轉(zhuǎn)基因苜蓿草KK179-2品系進行檢測。
[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】： 黃埔出入境檢驗檢疫局;廣東出入境檢驗檢疫局;
【基金】：廣東出入境檢驗檢疫局科技項目(2017GDK41、2016GDK60)
【分類號】：S541.9

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