Polycationic dendrimers were generated through simple and versatile reversible amine/aldehyde-imine chemistry. The inherent CD spectroscopic signal arising from the helical structures of single-stranded DNA (ssDNA) undergoes a dramatic amplification in the presence of the synthesised polycationic dendrimers. Compared to the first-generation core molecule, the second-generation dendrimer shows high spectroscopic responses upon chiral recognition of short ssDNA, owing to the combination of self-assembly and multivalency effects. The maximum signal variation is reached at the molar ratio at which the ratio between the negative charges in ssDNA balance the positive charges of the dendrimers, thus the approach enables differential recognition of ssDNAs of different lengths. Altogether, these results accelerate the simple and systematic discovery of efficient adaptive molecules for biomimetic recognition of ssDNA with high accuracy.