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Preparation and testing of E. coli S30 in vitro transcription translation extracts.

Methods in molecular biology (Clifton, N.J.) (2011-11-19)
James F Zawada
RÉSUMÉ

Crude cell-free extracts are useful tools for investigating biochemical phenomena and exploiting complex enzymatic processes such as protein synthesis. Extracts derived from E. coli have been used for over 50 years to study the mechanism of protein synthesis. In addition, these S30 extracts are commonly used as a laboratory tool for protein production. The preparation of S30 extract has been streamlined over the years and now it is a relatively simple process. The procedure described here includes some suggestions for extracts to be used for ribosome display.

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Pyruvate Kinase from rabbit muscle, Type VII, buffered aqueous glycerol solution, 350-600 units/mg protein