PURE (”Protein Synthesis Using Recombinant Elements”) is a defined cell-free system capable of transcribing mRNAs (using the T7 RNA polymerase) and translating proteins (using E. coli ribosomes and translation factors). PURE is built from individually purified components — proteins, RNAs, organic molecules, etc. The protocols here cover expressing and purifying protein components, preparing energy solutions and buffers, and assembling a complete PURE reaction.

• Getting Started
• Making PURE
• Make Protein Mix
• Prepare for Protein Purification
• Express and Purify Proteins
• Measure Prep Quality
• Assemble Protein Mix
• Energy Mix
• Assemble PURE Reactions