转基因水稻DNA样品浓度以及存放条件对PCR定性检测的影响

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摘要通过在不同模板DNA浓度（0.2~16 ng/μL）、不同转基因含量（0.05%~50%W/W ）以及不同模板存放温度（22℃、4℃和－20℃）和存放时间（1、2、3周和1，3个月）条件下进行转基因水稻外源成分的定性PCR检测，发现当模板浓度在0.4 ng/μL以上时所有引物均能扩增出目标片段，对于转基因含量为1.0%和0.1%的混合样品，能稳定检测出转基因成分所需的DNA最低浓度分别为1和2~4 ng/μL。模板DNA的不同温度存放条件对检测没有明显影响，长时间存放后PCR扩增产物条带亮度有所减弱。

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	王渭霞
	赖凤香
	洪利英
	傅强

关键词 ：转基因水稻, DNA浓度, PCR, 转基因检测

Abstract：The effects of DNA concentrations(0.2~16 ng/μL), transgene contents (50.0%~0.05%,W/W), storage temperatures(22℃，4℃，－20℃) and storage periods (1，2，3 weeks and 1, 3 months) on qualitative PCR detection of transgene from transgenic rice(Oryza sativa ssp.indica) were evaluated. The result showed that all the primers can amplified targeted fragment when DNA concentration was higher than 0.4 ng/μL. For mixed samples with 1.0% and 0.1% transgene, the lowest DNA concentration which was still detectable was 1 ng/μL and 2~4 ng/μL, respectively. Sample storage at the three tested different temperatures did not affect subsequent PCR detection. And the brightness of electrophoresis strips of PCR products were gradually weakened after 3 months storage of DNA sample.

Key words： Transgenic rice DNA concentration PCR Transgene detection

收稿日期: 2009-10-28

通讯作者: 傅强

引用本文:

王渭霞¹,赖凤香²,洪利英²,傅强³. 转基因水稻DNA样品浓度以及存放条件对PCR定性检测的影响[J]. , 2010, 18(5): 846-852.
, , ,. Effects of DNA Concentrations and Storage Conditions on PCR Detection of Transgenic Rice. , 2010, 18(5): 846-852.

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http://journal05.magtech.org.cn/Jwk_ny/CN/ 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2010/V18/I5/846