“
“Storage conditions are considered to be a critical component of DNA-based microbial community analysis methods. However, whether differences in short-term LDK378 in vivo sample storage conditions impact the assessment of bacterial community composition and diversity requires systematic and quantitative assessment. Therefore, we used barcoded pyrosequencing of bacterial 16S rRNA genes to survey communities, harvested from a variety of habitats [soil, human gut (feces)

and human skin] and subsequently stored at 20, 4, −20 and −80 °C for 3 and 14 days. Our results indicate that the phylogenetic structure and diversity of communities in individual samples were not significantly influenced by the storage temperature or the duration of storage. PD-0332991 purchase Likewise, the relative abundances of most taxa were largely unaffected by temperature even after 14 days of storage. Our results indicate that environmental factors and biases in molecular techniques likely confer greater amounts of variation to microbial communities than do differences in short-term storage conditions, including storage for up to 2 weeks at room temperature. These results suggest that many samples

collected and stored under field conditions without refrigeration may be useful for microbial community analyses. The treatment and 3-mercaptopyruvate sulfurtransferase handling of samples after collection is a critical aspect of a study design when using DNA-based methods

to compare the composition and diversity of microbial communities from environmental samples. It is widely assumed that microbial DNA must be extracted from the samples immediately after collection or, if this is not possible, that samples must be frozen (Rochelle et al., 1994). Samples stored at room temperature even for a short period before DNA is extracted are often considered unfit for downstream analyses because of changes to the microbial community. Although these assumptions are widespread, few and conflicting studies have directly tested the influence of storage conditions on DNA-based bacterial community analyses. For example, Dolfing et al. (2004) and Klammer et al. (2005) used DNA fingerprinting methods to show that the overall structure of soil bacterial communities was not strongly affected by storage conditions. Likewise, Roesch et al. (2009) reported only modest shifts in the bacterial diversity in only one of four human gut samples after 72 h of storage at room temperature. In contrast, both Tzeneva et al. (2009) and Ott et al. (2004) observed significant effects of storage conditions on the composition and diversity of microbial communities in soil and human gut samples, respectively. Nechvatal et al.