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Thorn, CE,Bergesch, C,Joyce, A,Sambrano, G,McDonnell, K,Brennan, F,Heyer, R,Benndorf, D,Abram, F
Molecular Ecology Resources
A robust, cost-effective method for DNA, RNA and protein co-extraction from soil, other complex microbiomes and pure cultures
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16S rRNA profiling DNA RNA and protein co-extraction metaproteomics microbial communities microbiomes pure cultures soil systems biology METAPROTEOME ANALYSIS EXTRACTION COMMUNITIES BACTERIAL RECOVERY IMPACT METATRANSCRIPTOMICS METAGENOMICS OPTIMIZATION INFORMATION
The soil microbiome is inherently complex with high biological diversity, and spatial heterogeneity typically occurring on the submillimetre scale. To study the microbial ecology of soils, and other microbiomes, biomolecules, that is, nucleic acids and proteins, must be efficiently and reliably co-recovered from the same biological samples. Commercial kits are currently available for the co-extraction of DNA, RNA and proteins but none has been developed for soil samples. We present a new protocol drawing on existing phenol-chloroform-based methods for nucleic acids co-extraction but incorporating targeted precipitation of proteins from the phenol phase. The protocol is cost-effective and robust, and easily implemented using reagents commonly available in laboratories. The method is estimated to be eight times cheaper than using disparate commercial kits for the isolation of DNA and/or RNA, and proteins, from soil. The method is effective, providing good quality biomolecules from a diverse range of soil types, with clay contents varying from 9.5% to 35.1%, which we successfully used for downstream, high-throughput gene sequencing and metaproteomics. Additionally, we demonstrate that the protocol can also be easily implemented for biomolecule co-extraction from other complex microbiome samples, including cattle slurry and microbial communities recovered from anaerobic bioreactors, as well as from Gram-positive and Gram-negative pure cultures.
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