Dataset Information

Comparison of preservation and storage methods for metaproteomics of intestinal microbiome samples

ABSTRACT: The aim of this study was to develop a suitable method to preserve fecal samples for metaproteomics analyses when flash-freezing is not an option. Fecal samples were collected from conventional adult C57BL/6 mice and combined into a fecal master mix. The fecal master mix was then split into 48 subsamples that were subjected to different preservation treatments. The following six preservation methods were tested: flash-freezing in liquid nitrogen followed by storage at -80°C, immersion in RNAlater® and storage at room temperature, immersion in RNAlater® and immediate storage at -80°C, immersion in 95% ethanol and storage at room temperature, immersion in a RNAlater-like buffer “NAP buffer” and storage at room temperature, and immersion in an autoclaved RNAlater-like buffer “Autoclaved NAP buffer” and storage at room temperature. Proteins were extracted from the samples after being stored for 1 and 4 weeks. There were 4 replicates per treatment and time-point. Samples were analyzed by LC-MS/MS and the data were analyzed with Proteome Discoverer against a large database of mouse microbiota protein sequences.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Mouse Gut Metagenome Mus Musculus (mouse)

SUBMITTER: Angie Mordant

LAB HEAD: Manuel Kleiner

PROVIDER: PXD024115 | Pride | 2021-11-25

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	AN1_140min_600ng_Run1.msf	Msf
	AN1_140min_600ng_Run1.pep.xml	Pepxml
	AN1_140min_600ng_Run1.raw	Raw
	AN2_140min_600ng_Run1.msf	Msf
	AN2_140min_600ng_Run1.pep.xml	Pepxml

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Publications

Evaluation of Sample Preservation and Storage Methods for Metaproteomics Analysis of Intestinal Microbiomes.

Mordant Angie A Kleiner Manuel M

Microbiology spectrum 20211215 3

A critical step in studies of the intestinal microbiome using meta-omics approaches is the preservation of samples before analysis. Preservation is essential for approaches that measure gene expression, such as metaproteomics, which is used to identify and quantify proteins in microbiomes. Intestinal microbiome samples are typically stored by flash-freezing and storage at -80°C, but some experimental setups do not allow for immediate freezing of samples. In this study, we evaluated methods to pr ...[more]

PMID: 34908431

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