| Steps 1–2, | Design of epegRNAs and nicking sgRNAs: 1 d |
| Step 3A, | Generation of epegRNAs or sgRNAs by Golden Gate cloning: 3 d |
| Step 3B, | Generation of epegRNAs or sgRNAs by isothermal assembly: 3 d |
| Step 3C, | Acquiring purified, chemically modified, synthetic epegRNAs, pegRNAs, or sgRNAs: 7–42 d |
| Steps 4–19, | Preparation of in vitro transcribed PEmax mRNA: 1–2 d |
| Step 20A, | Prime editing in HEK293T cells via plasmid transfection: 4–5 d |
| Step 20B, | Prime editing in primary human fibroblasts via RNA electroporation: 4–5 d |
| Steps 21–26, | Lysis of mammalian cells for HTS: 1 d |
| Steps 27–40, | HTS for prime editing analysis: 1–2 d |
| Steps 41–46, | HTS analysis: 1–4 h |
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