Extended Data Fig. 1: Optimization of the bridge sequence length.
From: Highly specific multiplexed RNA imaging in tissues with split-FISH
(a) Split probes were designed to target a polymorphic repeat region of the MUC5AC transcripts in A549 cell lines. Sample RNA FISH images of split bridge sequence length (x) ranging from 7 to 12 nucleotides (nt) in (b) unpaired and (c) paired split probes (orange and light blue sequences). Shorter (7–9 nucleotides) bridge lengths were able to suppress the binding of unpaired probes. However, using bridge lengths that were too short (7 + 7 nucleotides) resulted in poor binding even in paired probes. 9 + 9 nucleotides appeared to be the most optimal length. This experiment was repeated once with similar results.
