Vertex Pharmaceuticals 1 article published in JoVE Biology Rapid, Affordable, and Uncomplicated Production of Bacterial Cell-free Lysate Robert M. Cooper*1, Taishi Tonooka*1,2, Andriy Didovyk*1,3, Jeff Hasty1,4,5 1Biocircuits Institute, University of California, San Diego, 2Present address, Kyoto Institute of Technology, 3Present address, Vertex Pharmaceuticals, 4Department of Bioengineering, University of California, San Diego, 5Molecular Biology Section, University of California, San Diego This protocol describes a rapid and simple method to produce bacterial lysate for cell-free gene expression, using an engineered strain of Escherichia coli and requiring only standard laboratory equipment.