快速PCR方法在金银花真伪鉴别中的应用

Rapid PCR authentication Lonicera japanica

建立简单快速的金银花真伪分子鉴别方法。该研究依据金银花trn L-trn F 625位G/T SNP位点设计快速位点特异性PCR引物,优化位点特异性PCR条件,对金银花及其9种混淆品进行扩增及检测。当在87℃预变性1 min;87℃变性5s,68℃延伸5 s,30个循环时,通过加入SYBR Green I染料染色,金银花样品显绿色荧光,混淆品无荧光。整个PCR反应可在30 min内完成。该研究结果说明快速位点特异性PCR能简单快速鉴别金银花及其混淆品。

To simply and rapid authenticate Lonicera japanica. Rapid allele-specific PCR primer was designed base on trnL- trnF 625 G/T Single nucleotide polymorphism and the PCR reaction systems including annealing temperature was optimized; optimized results were performed to authenticate L. japanica and its 9 adulterants. When 100 x SYBR Green I was added in the PCR product of 87 ℃ initial denatured 1 min;87 ~C denatured 5 s,68 ℃ annealing 5 s,30 cycle; L. japanica visualize strong green fluorescence under 365 nm UV lamp whereas adulterants without. The results indicate rapid allele-specific PCR could authenticate L. japanica and its adulterants rapidly and simply.