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“Background Well-resourced culture collections Adenosine distribute bacteria mostly as freeze-dried ampoules [1, 2]. On the other hand, most research labs generally do not exchange lyophilized cultures and over the past 50 years a good proportion of bacterial exchanges were either in

agar stabs or on impregnated glycerolized discs, as also used by the Coli Genetic Stock Center (CGSC). Generally, comparison of storage and shipping conditions test for viability and all of the above methods work well in this regard for Escherichia coli. Recently however, we became concerned about heterogeneity arising during storage and exchange of cultures for two reasons. Firstly, our recent studies with the ECOR collection [3] indicated a number of phenotypes had changed from those reported earlier (unpublished results). Others have also noted discrepancies in results with the ECOR collection between laboratories [4]. Secondly, in recently exchanged stock cultures of E. coli K-12 between the Ferenci and Spira laboratories, we noted heterogeneities in some of the phenotypes we routinely assay. In this communication, we investigated the source of this heterogeneity and the role of storage conditions during shippage. The instability of cultures and possible heterogeneities have been noted in several settings. Bacteria in long term stab cultures were found to change in a number of respects [5–8].