Lab Manual
biological reagents
Cell Free Systems
In Vivo Systems
misc methods
CELL FREE SYSTEMS > General Cell Free Translation  

> Reticulocyte Lysate and Wheat Germ Extracts

  > tRNA Crosslinking  

> Immunoprecipitation of translation products

  > Translation in RRL  

> History and evolution of cell-free protein synthesizing systems


> Import Into Mitochondria


> Wheat Germ Cell-Free Translation

  > E coli Translation

> Translation Protocol


> Sample Protocol


> Troubleshooting


> Lysate Preparation of S-30


> Membrane-Free Ribosomes (mfrs)


> Ribosomal Pellet


> Dialysis Tubing Work-up


> Membrane-Free Translation System

    > Buffers and Reagents



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The S-30 system for E. coli coupled transcription-translation was first described by Zubay. Proteins are synthesized from an exogenous DNA template. The yield from this system is comparable to that of RRL, and the ease of use is somewhat more simple. One must remember that a bacterial promoter (i.e. not SP6 or T7) and Shine-Delgano sequence must be present for translation of the desired protein.

As prepared, the S-30 system contains endogenous bacterial membrane vesicles. These can be removed by high speed centrifugation, but membrane-free ribosomes must be isolated and added back to this system in order to restore translation.