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基于下一代测序的CRISPR/Cas9基因组编辑油菜种子分析的可重复性

Steffen Pallarz¹, Stefan Fiedler², Daniela Wahler¹

¹Department Genetic Engineering and Other Biotechnological Processes, Federal Office of Consumer Protection and Food Safety (BVL), P.O. Box 110260, 10832 Berlin, Germany.

Food chemistry. Molecular sciences

|October 12, 2023

概括

下一代测序 (NGS) 显示了在实验室中检测食品和料中的转基因生物 (GMO) 的高可重现性. 长读全基因组测序 (WGS) 是有希望的,但由于变异性,对转基因检测尚未成为常规.

科学领域:

农业生物技术农业生物技术
分子生物学分子生物学
遗传学遗传学是一个

背景情况:

下一代测序 (NGS) 对于在食品,料和种子中检测转基因生物 (GMO) 是至关重要的.
标准化转基因检测工作流程需要可靠的实验室内和实验室间可重复性.
评估不同的NGS方法对于强大的转基因生物分析至关重要.

研究的目的:

评估短读和长读目标NGS和长读全基因组测序 (WGS) 的可复制性,以检测转基因生物.
为了比较三个独立实验室不同测序策略的性能.
确定各种NGS方法在执法环境中适用于常规GMO检测的适用性.

主要方法:

使用目标NGS (短读和长读) 和WGS (长读) 的复制样本的比较分析.
样本包括基因组编辑的油菜 (OSR),入商品OSR和父系叶片材料.
在三个独立实验室中,数据分析了可复制性,质量和映射效率.

主要成果:

针对性的NGS (短读和长读) 在所有实验室都表现出高的可重复性和质量.
与转基因相关的序列检测在使用向NGS的设施中是一致的.
长期阅读的WGS为复杂的基因组提供了卓越的映射,但显示了显著的实验室间变异.
长读WGS目前不适合在执法实验室的常规使用.

结论:

向NGS是一种可靠和可重复的方法,用于在不同实验室中检测食品和料中的转基因生物.
长读测序为绘制复杂基因组提供了优势,但需要进一步标准化以进行常规的转基因检测.
长期读取的WGS可复制性的当前限制阻碍了其在监管执法环境中立即采用.

关键词:

检测检测检测检测检测基因编辑 (GE) 是指基因编辑.转基因生物 (转基因生物) 是指转基因生物.国家存款系统数据质量.NGS的可复制性NGS的可复制性新基因组技术 (NGT) 是指新的基因组技术.

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