Double sided adhesive

Double sided adhesive (3M, Maplewood, MN) with 1.5 mm diameter holes was applied to the surface of a wicking layer (ShamWow^®, Wallingford, CT). 2 mm disks of functionalized Fusion 5 filters were carefully adhered to the wicking layer, centered over the 1.5 mm holes in the adhesive. 1 mL of 1x PBS (pH 5) with varying concentrations of synthetic proviral DNA target was applied to the filters using a 1 mL syringe. The solution was wicked through the filters into the bottom layer (Figure S3).
The filters were then moved to Double sided adhesive on the surface of custom 3D printed rings (Figure 1c, black rings) and dried for 10 minutes in the custom drying chambers (Figure 1b, blue component). H1 and H2 conjugated with Cy3 and Cy5, respectively, were diluted to 5 nM concentrations in 0.5X SSC with 10 mM MgCl₂, 0.05% Tween 20, and 6% PEG (brought to pH 8 using Tris base) and were applied to the paper disks as 5 μL droplets. The PEG concentration was increased in these experiments to 6% due to further optimization of the crowding buffer concentrations. The reactions were fully evaporated for 60 minutes in the custom drying chambers and processed using the fluorescence microscope and FRET cube as described earlier with the exception that the images were taken at a gain of 4 and 500 ms exposure and images were converted to RGB color prior to analysis.

Double sided adhesive (3M, Maplewood, MN) with 1.5 mm diameter holes was applied to the surface of a wicking layer (ShamWow^®, Wallingford, CT). 2 mm disks of functionalized Fusion 5 filters were carefully adhered to the wicking layer, centered over the 1.5 mm holes in the adhesive. 1 mL of 1x PBS (pH 5) with varying concentrations of synthetic proviral DNA target was applied to the filters using a 1 mL syringe. The solution was wicked through the filters into the bottom layer (Figure S3).
The filters were then moved to Double sided adhesive on the surface of custom 3D printed rings (Figure 1c, black rings) and dried for 10 minutes in the custom drying chambers (Figure 1b, blue component). H1 and H2 conjugated with Cy3 and Cy5, respectively, were diluted to 5 nM concentrations in 0.5X SSC with 10 mM MgCl₂, 0.05% Tween 20, and 6% PEG (brought to pH 8 using Tris base) and were applied to the paper disks as 5 μL droplets. The PEG concentration was increased in these experiments to 6% due to further optimization of the crowding buffer concentrations. The reactions were fully evaporated for 60 minutes in the custom drying chambers and processed using the fluorescence microscope and FRET cube as described earlier with the exception that the images were taken at a gain of 4 and 500 ms exposure and images were converted to RGB color prior to analysis.