Figure 3
Characterization of the learning protocol on the basis of the ethidium bromide staining denatured Urea polyacrylamide gel electrophoresis (Urea-PAGE) at 60°C. (a) Digested E. coli gDNA with 4-12% Urea-PAGE. The lanes represent: (1) molecular size marker for single-stranded (ss) DNA of 100 to 1000 bases in 100-base increments and (2) digested E. coli gDNA (avg. ~200 bases). (b) Learning products with 4-20% Urea-PAGE. The lanes represent: (1) molecular size marker for ssDNA from 100 base in 100-base increments, (2) molecular size marker for ssDNA of 40 to 100 bases in 10-base increments, (3) memory tags (40 bases), (4) learning reaction mixture (i.e., memory tag + digested gDNA) after annealing and digestion (A → D) (5) digested gDNA only after annealing and extension (A → E), (6) memory tags only after A → E, (7) learning reaction mixture after A → E, (8) digested gDNA only after annealing, extension, and digestion (A → E → D), (9) memory tags only after A → E → D, and (10) learning reaction mixture after A → E → D. The learning protocols were implemented with 5′-amine modified memory tags and the product of each run was separated by the magnetic bead separation. LPH is the high-molecular weight (>80 bases) learned products and LPS relatively short (50 – 80 bases) learned products.