“In Vitro Construction and Long Read Sequencing Analysis of a 24 Kb Long Artificial DNA Sequence Encoding the Universal Declaration of the Rights of Man and of the Citizen ”
biorxiv(2023)
摘要
In absence of 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. We propose in this report a fully in vitro protocol to generate very long double-stranded DNA molecule starting from commercially available short DNA blocks in less than 3 days. This innovative application of Golden Gate assembly allowed us to streamline the assembly process to produce a 24 kb long DNA molecule storing part of the Universal Declaration of Human rights and citizens. The DNA molecule produced can be readily cloned into suitable host/vector system for amplification and selection.### Competing Interest StatementThe authors have declared no competing interest.
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