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 3days usingGolden Gate assembly. This innovative applicationallowed us to streamline the process to produce a 24kb-long DNA molecule storing part of theDeclaration 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.
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