CIIM Abstract

Curr. Issues Intestinal Microbiol. (2006) 7: 29-34

Freezing at -80 Degrees Distorts the DNA Composition of Bacterial Communities in Intestinal Samples

Lars Molbak, Helle M. Sommer, Kaare Johnsen, Mette Boye, Markku Johansen, Kristian Moller and Thomas D. Leser

Terminal-restriction fragment length polymorphism (TRFLP) was used to evaluate how to store intestinal specimens for bacterial community analysis. Bacterial communities are increasingly often described by means of DNA-based methods and it is common practice to store intestinal or faecal specimens either at -20 degrees C or -80 degrees C. In this study, samples of intestines from five different pigs were stored at -80 degrees C and -20 degrees C, respectively and a thawing and freezing procedure was carried out three times for each intestinal per pig per temperature. The cumulative sum of the T-RFLP peak heights (T-RF intensities) decreased as the temperature decreased. The composition of the bacterial community changed when stored at -80 degrees C compared to the samples stored at -20 degrees C. Thus it is recommended from this study that samples of intestinal content are stored at -20 degrees C before use for bacterial community analysis, instead of the current practice at -80 degrees C.

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