Abstract
<jats:p>This paper does not present new experimental data, but it offers unique insights into the secondary structure of DNA and its replication based on a non-canonical structural model herein referred to as the ambidextrous double-helix model, in which the two strands are not strictly right-hand wound. Analysis of positively and negatively supercoiled plasmids revealed that the differences between them could be reasonably explained by the ambidextrous double-helix model, but not by the classical double helix model. The superhelical structure of DNA has been understood and explained in an unprecedented way, which may help us better unravel the mysteries of nature. A framework operating at the subconscious level—constructed through years of rigorous scrutiny, profound introspection, and the cross-verification of extensive evidence from many different laboratories—has given rise to a series of concepts in the field of DNA topology; these theoretical constructs, rooted in direct experimental observations, are highly likely to offer peers in the field fresh perspectives and new avenues for validation.</jats:p>