Fig. 6: Fluorescence-assisted sorting of proteinosomes for selective file retrieval.
From: DNA storage in thermoresponsive microcapsules for repeated random multiplexed data access
a, Schematic of membrane- and localized DNA-based barcoding of proteinosomes. The proteinosome membranes are labelled with either DyLight 405 (blue) or FITC (green); additionally, fluorescent biotinylated ssDNA labelled with Cy3 (orange; F4) or Cy5 (red; F5) can be localized inside the proteinosomes alongside the DNA files. b, Individual, barcoded populations of localized files can be pooled together in a single searchable library. Proteinosomes are sorted via FACS based on the four barcodes (Methods). Effective sorting was verified using qPCR. c, FACS dot plots of FITC versus DyLight 405 (left) and Cy3 versus Cy5 (right) labels. Both plots contain two distinct clusters that were used to sort the pooled proteinosomes into four populations. Supplementary Fig. 16 shows the full gating strategy and histograms of individual fluorescent channels. Sequences for F4 and F5 are listed in Supplementary Table 5. d, Sorting selectivity as determined by qPCR. The horizontal lines indicate the mean file fractions; the circles indicate three individual data points. Unintentionally mis-sorted files account for 8% of the total DNA concentration, whereas the intended file sorting accounts for 75% of the total. There is a statistically significant difference of 8.4-fold change in the relative fractions observed using a two-sided Welch’s t-test (p = 0.00096). Sequences for F4, F5 and the primers used are listed in Supplementary Table 6.
