Mølbak Lars, Sommer Helle M, Johnsen Kaare, Boye Mette, Johansen Markku, Møller Kristian, Leser Thomas D
Danish Institute for Food and Veterinary Research, Copenhagen.
Curr Issues Intest Microbiol. 2006 Mar;7(1):29-34.
Terminal-restriction fragment length polymorphism (T-RFLP) 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.
末端限制性片段长度多态性(T-RFLP)被用于评估如何储存肠道标本以进行细菌群落分析。越来越多地通过基于DNA的方法来描述细菌群落,并且将肠道或粪便标本储存在-20℃或-80℃是常见的做法。在本研究中,分别将来自五头不同猪的肠道样本储存在-80℃和-20℃,并且对每头猪的每个肠道样本在每个温度下进行三次解冻和冷冻程序。随着温度降低,T-RFLP峰高(T-RF强度)的累积总和下降。与储存在-20℃的样本相比,储存在-80℃时细菌群落的组成发生了变化。因此,本研究建议在用于细菌群落分析之前,将肠道内容物样本储存在-20℃,而不是目前-80℃的做法。