University of Oxford, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, Oxford OX3 9DS, UK.
STAR Protoc. 2021 May 5;2(2):100507. doi: 10.1016/j.xpro.2021.100507. eCollection 2021 Jun 18.
Here, we describe a highly efficient, medium-throughput strategy for cloning and screening of putative enhancers using the chick embryo. By incorporating 48 unique nanotags for use in NanoString nCounter® across three different fluorescent reporters and developing a rapid and efficient digestion/ligation type IIs restriction enzyme-based cloning protocol, we develop a multiplexed approach for rapidly identifying enhancer activity. For complete details on the use and execution of this protocol, please see Williams et al. (2019).
在这里，我们描述了一种高效、中通量的策略，用于使用鸡胚克隆和筛选假定的增强子。通过在三个不同的荧光报告基因中结合 48 个独特的纳米标签用于 NanoString nCounter®，并开发一种快速有效的消化/连接 IIs 型限制性内切酶克隆协议，我们开发了一种用于快速鉴定增强子活性的多路方法。有关该协议的使用和执行的完整详细信息，请参见 Williams 等人（2019 年）。
