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.
摘要：
在没有DNA模板的情况下,预定义序列的长双链DNA分子的从头算产生尤其具有挑战性。DNA合成步骤仍然是许多应用的瓶颈，例如祖先基因功能评估，分析选择性剪接或基于DNA的数据存储。在本报告中，我们提出了一种完全体外的方案，以使用GoldenGate组装在不到3天的时间内从市售的短DNA块开始产生非常长的双链DNA分子。这种创新的应用使我们能够简化生产24kb长的DNA分子的过程，该分子存储了1789年《人权宣言》和《公民权利宣言》的一部分。产生的DNA分子可以容易地克隆到合适的宿主/载体系统中用于扩增和选择。
