%0 Journal Article
%T Fully in vitro iterative construction of a 24 kb-long artificial DNA sequence to store digital information.
%A Leblanc J
%A Boulle O
%A Roux E
%A Nicolas J
%A Lavenier D
%A Audic Y
%J Biotechniques
%V 76
%N 5
%D 2024 May 4
%M 38573592
%F 2.746
%R 10.2144/btn-2023-0109
%X In the absence of a DNA template, the ab initio production of long double-stranded DNA molecules of predefined sequences is particularly challenging. The DNA synthesis step remains a bottleneck for many applications such as functional assessment of ancestral genes, analysis of alternative splicing or DNA-based data storage. In this report we propose a fully in vitro protocol to generate very long double-stranded DNA molecules starting from commercially available short DNA blocks in less than 3 days using Golden Gate assembly. This innovative application allowed us to streamline the process to produce a 24 kb-long DNA molecule storing part of the Declaration of the Rights of Man and of the Citizen of 1789 . The DNA molecule produced can be readily cloned into a suitable host/vector system for amplification and selection.