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Author:
Date:
2017-11-20
[Abstract] This protocol describes how to order and directly assemble uracil-containing non-clonal DNA fragments by uracil excision based cloning (USER cloning). The protocol was generated with the goal of making synthesized non-clonal DNA fragments directly compatible with USERTM cloning. The protocol is highly efficient and would be compatible with uracil-containing non-clonal DNA fragments obtained from any synthesizing company. The protocol drastically reduces time and handling between receiving the synthesized DNA fragments and transforming with vector and DNA fragment(s).
[摘要] 该协议描述如何通过基于尿嘧啶切除的克隆（USER克隆）命令并直接组装尿嘧啶非克隆DNA片段。 该方案的目的是制备与USER TM克隆直接兼容的合成非克隆DNA片段。 该协议是高效率，并将与从任何合成公司获得尿嘧啶非克隆DNA片段相容。 该方案大大减少了接收合成的DNA片段和用载体和DNA片段转化之间的时间和处理。
【背景】对于合成的DNA，非克隆线性DNA片段（NCDF）已经成为更廉价和更快速的替代克隆片段的方法，并且在循环载体中被测序。 NCDF可以被认为是IKEA DNA合成的解决方案，在这个解决方案中，客户将他们的DNA片段组装成一个选择的载体，随后必须验证最终构建体的序列。在这个协议中，我们从Thermo Fisher Scientific ...
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