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Schematic representation of the in vitro transcription (IVT)
The DNA template generated (step 5) contains the T7 promoter followed by 5′GG and the processed direct repeat for RfxCas13d gRNA and the specific binding sequence of 22 nt. The overnight IVT product is the sgRNA for RfxCas13d system with the tail (in blue) and 22 nt of binding site to anneal with the mRNA target.
