Evaluating methods for the preservation and extraction of DNA and RNA for analysis of microbial communities in marine sponges

摘要: Abstract Preservation and extraction of high quality DNA RNA from biological samples are becoming increasingly important for genomics, transcriptomics microbial community analyses. Of major recent interest, due to their ecological biotechnological importance, the communities microorganisms associated with marine sponges. In this study, we systematically evaluated protocols preservation (RNA later , liquid nitrogen, lyophilized, frozen) (CTAB-based extraction, isolation via TRIzol two co-extraction methods) nucleic acids using New Zealand high-microbial-abundance sponge Ancorina alata . The quantity were assessed spectrophotometry, agarose gel electrophoresis microcapillary electrophoresis. Although all resulted in sufficiently and/or downstream applications, there significant differences yield extracted. All methods maintained mRNA sufficient integrity amplify prokaryotic glutamine synthetase gene. Denaturing gradient (DGGE) analysis 16S rRNA gene- rRNA-derived fragments revealed several unique bands profiles. However, no changes attributable either or method. Optimized successfully tested on three other Our results suggest that whilst choice method does influence isolated sponges, performed favorably, therefore protocol can be made based practical considerations including ease use, time availability cost.