School of Life Sciences, Zhengzhou University, No. 100 Science Avenue, Zhengzhou, Henan 450001, China.
Institute of Logistics Science and Engineering, Shanghai Maritime University, 1550 Haigang Avenue, Pudong District, Shanghai 201306, China.
J Microbiol Methods. 2022 Nov;202:106565. doi: 10.1016/j.mimet.2022.106565. Epub 2022 Sep 9.
Cell lysates from a laboratory strain of Escherichia coli can be exploited for seamless DNA cloning in vitro, which is named the seamless ligation cloning extract (SLiCE) cloning method. The SLiCE method can incorporate DNA fragments into a vector to achieve conventional DNA cloning and is more cost-effective than commercially seamless DNA cloning kits. In this study, we found that the SLiCE extracts could easily be prepared with different methods, such as 3% Triton X-100 lysis buffer, 3% SDS lysis buffer, or freeze-thaw cycles. At high E. coli transformation efficiency, the SLiCE extracts prepared using different simple and ultra-low cost methods did not affect the DNA cloning efficiency. These results further revealed that the SLiCE cloning method can be efficiently used for seamless DNA cloning in vitro.
实验室大肠杆菌菌株的细胞裂解物可用于体外无缝 DNA 克隆，该方法被命名为无缝连接克隆提取物（SLiCE）克隆方法。SLiCE 方法可将 DNA 片段整合到载体中，实现常规 DNA 克隆，并且比商业无缝 DNA 克隆试剂盒更具成本效益。在本研究中，我们发现 SLiCE 提取物可以使用不同的方法轻松制备，例如 3% Triton X-100 裂解缓冲液、3% SDS 裂解缓冲液或冻融循环。在高大肠杆菌转化效率下，使用不同简单和超低成本方法制备的 SLiCE 提取物不会影响 DNA 克隆效率。这些结果进一步表明，SLiCE 克隆方法可有效地用于体外无缝 DNA 克隆。
