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用荧光PCR毛细管电泳技术在基因型CRISPR / Cas9介导的敲除突变体的高通量形式

JoVE Journal
Genética

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JoVE Journal Genética

Using a Fluorescent PCR-capillary Gel Electrophoresis Technique to Genotype CRISPR/Cas9-mediated Knockout Mutants in a High-throughput Format

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用荧光PCR毛细管电泳技术在基因型CRISPR / Cas9介导的敲除突变体的高通量形式

DOI:

10.3791/55586-v

•

08:25 min

•

April 08, 2017

•

Muhammad Khairul Ramlee, Jing Wang, Alice M. S. Cheung, Shang Li

¹Cancer & Stem Cell Biology Programme,Duke-NUS Medical School

Capítulos

00:05Título
00:55Sample Preparation and Amplification of CRISPER/Cas9 Regions with Fluorescent PCR
04:32Capillary Gel Electrophoresis and Electropherogram Analysis
06:25Results: Genotyping HEPG2 Targeted with CRISPER/Cas9 Construct Against NAP1L1
07:27Conclusion

Summary

Tadução automática

这里所描述的基因分型技术，荧光聚合酶链式反应（PCR），以毛细管凝胶电泳，其耦合，允许核酸酶介导敲除克隆的高通量基因分型。它规避所面临的其他基因分型技术的限制，并且比测序方法更具成本效益。

Tags

CRISPR/Cas9 Knockout Mutants Genotyping Fluorescent PCR Capillary Gel Electrophoresis High-throughput Cell Culture Cloning Cell Lysis PCR

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