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例如,为了创建有成本效益的NGS文库,可以使用所述CRISPR系统破坏靶基因的编码序列,并且可以通过下一代测序(例如,在离子激流(Ion Torrent)PGM系统上)同时筛选经所述CRISPR相关蛋白转染的克隆。关于如何制备NGS文库的详细描述可见于例如Bell等人,“Ahigh-throughputscreening strategy for detecting CRISPR-Cas9induced mutations...
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High-throughput sequencing was then performed using an Ion Torrent S5 sequencer (Thermo Fisher Scientific). The raw data were filtered to remove low-quality reads using FASTP (Chen et al., 2018). Filtered reads were normalized using BBNorm software (Biowulf, USA) and then compared with the ...
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