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Fig. 1 | Cell & Bioscience

Fig. 1

From: Customized one-step preparation of sgRNA transcription templates via overlapping PCR Using short primers and its application in vitro and in vivo gene editing

Fig. 1

A schematic diagram of the conventional methods to generate sgRNA in vitro. a The simple method is direct sgRNA chemical synthesis. The conventional methods to generate sgRNAare in vitro transcription using templates. b The direct method to prepare the templates is to DNA synthesize T7-sgRNA fragments as transcription templates. c One of methods is to construct a recombinant plasmid with T7-sgRNA backbone sequence to be taken as the sgRNA transcription template. d Another method is to take plasmids with T7-sgRNA backbone as PCR templates to amplify transcription templates through using a forward long primer and a reverse primer

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