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. 2022 Nov 8;3(4):101829. doi: 10.1016/j.xpro.2022.101829

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© 2022 The Author(s)

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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DNA sequences of BCS components

(A) The foundation consists of a 16-nt foundation base, 8-nt foundation barcode, and 7-nt bridge-binding sequence. The target tail consists of a 5′ phosphate (represented as “/5Phos/”), 20-nt target base sequence, and a 21- or 25-nt spacer. Red bars symbolize covalent linkage to the target. The forward cololinker consists of 16-nt foundation base complement, 20-nt P7 complement, 44-nt T spacer, and 20-nt hybridization region. The reverse cololinker consists of a 20-nt target base complement, 20-nt P7 complement, 44-nt T spacer, and 20-nt hybridization region.

(B) The binder tail consists of a 9-nt ligation spacer, 12-nt binder barcode, 24-nt restriction site spacer (containing an EcoR1 site), and 15-nt T-spacer. Red bars indicate covalent linkage to the nanobody binder. The universal bridge consists of a 5′ region complementary to the restriction site spacer (containing the EcoRI restriction site), a region of 12 nucleotides of 5-nitroindole universal bases (represented as “5”) that can hybridize with any binder barcode, a ligation spacer complement, and a 3′ overhang complementary to the 5′ region of all foundations or previously ligated binder barcode sequences.