Journal: bioRxiv

Article Title: Fuel-driven catalytic molecular templating

doi: 10.64898/2026.02.18.706517

Figure Lengend Snippet: (a) Schematic of binding of S 1 to T 11 via TMSD in Step 1 of the catalytic cycle, and the concentration of S 1 -T 11 against time when reacting 10 nM of S 1 -L with varying amounts of T 11 . (b) Schematic of the binding of R 1 to S 1 via HMSD in Step 2 of the catalytic cycle, and the concentration of S 1 -T 11 -R 1 against time when reacting 10 nM of S 1 -T 11 with varying amounts of R 1 . (c) Fuel-driven release of the template T 11 to form S 1 -R 1 -D 7 in Step 3 of the catalytic cycle, and concentration of S 1 -R 1 -D 7 against time when reacting 10 nM of S 1 -T 11 -R 1 with varying concentrations of D 7 . Details of how concentrations are inferred from raw fluorescence traces are provided in sections SI.3 and SI.4 of the Supporting Information, and the data are obtained from three replicas with the mean and range plotted in each case. All quoted reactant concentrations are those intended in the experimental design; pipetting errors and uncertainties in the supplied stock concentration lead to variation in the true levels, which are estimated as discussed in sections SI.3.3 - SI.3.5.

Article Snippet: Then the gel was imaged on an Amersham Typhoon fluorescence gel scanner using the following instrument settings: blue (excitation: 488 nm, emission: 525/20 nm, photomultiplier tube voltage: 344 V), and green (excitation: 532 nm, emission: 570/20 nm, photomultiplier tube voltage: 343 V. The pseudocolour cropped image reported in and the full image reported in Supporting Figure S13 are generated by merging the original greyscale images using ImageJ software, after adjusting the brightness for better visibility and reduced background.

Techniques: Binding Assay, Concentration Assay, Fluorescence

Journal: bioRxiv

Article Title: Fuel-driven catalytic molecular templating

doi: 10.64898/2026.02.18.706517

Figure Lengend Snippet: (a)-(d) Fluorescence relative to positive control of 10 nM S 1 -R 1 -D, measured in experiments in which varying concentrations of template T 11 were added to solutions of 10 nM each of S 1 -L, R 1 and D j , in which j = 5, 6, 7, 10 indicates | ith |. Curves are fit with as outlined in section 4.2.1 of the Supporting Information. (e) Catalytic turnover with excess concentration of fuel D 6 . Fluorescence relative to a 10 nM S 1 -R 1 -D 6 positive control, when 2 nM of T 11 was added to a solution of 10 nM S 1 -L, 10 nM R 1 alongside excess concentrations (10 - 50 nM) of D 6 . (f) Catalytic turnover with a very high concentration of monomers compared to the template. Fluorescence relative to a 50 nM S 1 -R 1 -D 7 positive control after T 11 at a range of low concentrations was injected into a solution of 50 nM S 1 -L, 50 nM R 1 and 100 nM D 7 . (a)-(e) are data obtained from three replicas with the mean and range plotted in each case, whereas f is from data repeated only once; a third experiment showed a technical malfunction and is reported separately along with data at longer times in Supporting Figure S10.

Article Snippet: Then the gel was imaged on an Amersham Typhoon fluorescence gel scanner using the following instrument settings: blue (excitation: 488 nm, emission: 525/20 nm, photomultiplier tube voltage: 344 V), and green (excitation: 532 nm, emission: 570/20 nm, photomultiplier tube voltage: 343 V. The pseudocolour cropped image reported in and the full image reported in Supporting Figure S13 are generated by merging the original greyscale images using ImageJ software, after adjusting the brightness for better visibility and reduced background.

Techniques: Fluorescence, Positive Control, Concentration Assay, Injection

Journal: bioRxiv

Article Title: Fuel-driven catalytic molecular templating

doi: 10.64898/2026.02.18.706517

Figure Lengend Snippet: (a) Fluorescence relative to a positive control of 10 nM S 1 -R 1 -D when 2 nM of template (either T 11 , T 12 , T 21 or T 22 ) was added to 10 nM each of S 1 -L, R 1 and D. Flourescence increases as S 1 was released from its duplex with quencher-labelled L. (b) Fluorescence in the blue channel observed after adding 5 nM of each template to a mixture of all four monomers and D, each at 10 nM, relative to positive control of 10 nM S 1 -R 2 -D (also shown is a second control of 10 nM S 1 -R 2 -D). (c) Fluorescence in the green channel observed in the same experiments as (b), relative to positive control of 10 nM S 2 -R 2 -D (also shown is 10 nM S 2 -R 1 -D). (d) Schematic illustrating the information propagation experiment from (b) and (c), in which individual templates were added to monomer pools containing of all monomers and fuel to selectively produce a matching product as in plots. (e) Gel electrophoresis image supporting the specificity of the reactions. The untrimmed gel data is reported in Supporting Figure S13.

Article Snippet: Then the gel was imaged on an Amersham Typhoon fluorescence gel scanner using the following instrument settings: blue (excitation: 488 nm, emission: 525/20 nm, photomultiplier tube voltage: 344 V), and green (excitation: 532 nm, emission: 570/20 nm, photomultiplier tube voltage: 343 V. The pseudocolour cropped image reported in and the full image reported in Supporting Figure S13 are generated by merging the original greyscale images using ImageJ software, after adjusting the brightness for better visibility and reduced background.

Techniques: Fluorescence, Positive Control, Control, Nucleic Acid Electrophoresis