Supplementary Fig. 3: Quality control for in vitro transcribed RNA.
From: GOTI, a method to identify genome-wide off-target effects of genome editing in mouse embryos
a–d, Preparation of in vitro transcription templates for SpCas9 (a), BE3 (b), Cre (c) and sgRNA (d). e, Amount and quality check for SpCas9, BE3, Cre and sgRNA transcripts. f, Failed in vitro transcription of spCas9 mRNA. For a successful in vitro transcription, a major band of interested transcript should appear like in (e), or it will be a failed reaction (f). In gel running results, smear could often be found around the major band of tailed transcripts, and size changes of transcripts might appear for RNA folding. The gene editing ability of these transcripts would not be changed as long as the major band could be clearly distinguished. DNA and RNA were run on 1% denaturing agarose stained with nucleic acid dye.
