Fig. 1From: Customized one-step preparation of sgRNA transcription templates via overlapping PCR Using short primers and its application in vitro and in vivo gene editingA 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 primerBack to article page