Fig. 3: Enzymatic amplification of DNA localized inside proteinosomes.
From: DNA storage in thermoresponsive microcapsules for repeated random multiplexed data access
a, Experimental design to measure the enzymatic amplification of localized DNA templates inside proteinosomes containing 4 µM Tamavidin 2-HOT. DNA strands with (178 bp; A1T1) or without (168 bp; U1T1S) a biotin end modification were added to the proteinosomes and removed with five washing steps (Methods). An amplification reaction mixture containing enzymes, primers, dNTPs and dsDNA-specific EvaGreen dye was then added and the amplification of DNA localized in the proteinosomes was measured using real-time fluorescence monitoring. States of high (T < LCST) and low (T > LCST) membrane permeability are shown as thin or thick blue dashed circles, respectively. b, qPCR results of amplified DNA from proteinosomes incubated with either biotin-labelled DNA or non-labelled DNA (Methods). The horizontal lines indicate the mean threshold cycle (Ct) for three experiments; the points represent individual experiments. A statistically significant difference of 4.4 cycles between the two conditions is observed using a two-sided Welch’s t-test (p = 0.049). Individual amplification traces are shown in Supplementary Fig. 11. The sequences used are listed in Supplementary Table 2. c, SDA results of isothermally amplifying DNA from proteinosomes incubated with biotin-labelled DNA or non-labelled DNA (Methods). The horizontal lines denote the mean production rate for three experiments; the points represent individual experiments. A statistically significant 8.6-fold difference in rates is observed between the two conditions using a two-sided Welch’s t-test (p = 0.004). Individual amplification traces are shown in Supplementary Fig. 12. The sequences used are listed in Supplementary Table 2.
