Ligation-independent cloning (LIC) is a simple, rapid, and efficient method for high-throughput cloning. In this system, linear plasmid vector and insert DNA are treated to generate complementary single-stranded overhangs that anneal during a short incubation. The LIC system is adaptable for use with any vector following an alteration of the vector sequence. This chapter describes the creation of an LIC-compatible vector, with tips on how to make any vector LIC-enabled. It also includes a protocol for generating high-quality linearized vector template for the LIC reaction. Lastly, a step-by-step protocol of the LIC reaction is outlined, with useful tips and tricks for optimization and screening.
CITATION STYLE
Doyle, S. A. (2005). High-throughput cloning for proteomics research. Methods in Molecular Biology (Clifton, N.J.), 310, 107–113. https://doi.org/10.1007/978-1-59259-948-6_7
Mendeley helps you to discover research relevant for your work.