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 teach you how to express and purify proteins, prepare energy solutions and buffers, and assemble a complete PURE reaction.
Make Protein Purification Buffers and Media
Grow and Induce Expression Strains
Purify Proteins by Ni2+ Gravity Column
Exchange Buffers and Concentrate by Spin Filtration