Growing and Handling of Bacterial Cultures within a Shared Core Facility for Integrated Structural Biology Program

Abstract

We have established and optimized standard operating procedures for growing and handling bacterial cultures in a shared core laboratory to support Integrative Structural Biology. The Integrative Structural Biology effort within the Biomolecular Research Center allows researchers to generate new knowledge about protein and RNA structure and function. We aim to understand how biomolecules assemble into stable structures and how structural dynamics impacts their function. Here we describe specific procedures for growing and handling bacterial cultures for overexpression and isolation of recombinant proteins, 15 N/ 13 C uniform labeling of recombinant proteins, protein isolation and purification, and analysis of protein solubility that are ideal for implementation in a shared research core laboratory that serves a multitude of diverse customers and research laboratories. pipettor speeds up the aliquoting process minimize the time that the competent cells are manipulated, their competency. competency of ca. 10 7 –10 8 CFU/ μ g of plasmid DNA.