{Reference Type}: Journal Article
{Title}: Fully in vitro iterative construction of a 24 kb-long artificial DNA sequence to store digital information.
{Author}: Leblanc J;Boulle O;Roux E;Nicolas J;Lavenier D;Audic Y;
{Journal}: Biotechniques
{Volume}: 76
{Issue}: 5
{Year}: 2024 May 4
{Factor}: 2.746
{DOI}: 10.2144/btn-2023-0109
{Abstract}: 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.