A single microfluidic device for multi-omics analysis sample preparation

被引:0
|
作者
Kumar, Ranjith Kumar Ravi [1 ]
Haddad, Iman [1 ]
Ndiaye, Massamba Mbacke [1 ]
Marbouty, Martial [2 ]
Vinh, Joelle [1 ]
Verdier, Yann [1 ]
机构
[1] PSL Univ, Spectrometrie Masse Biol & Prote SMBP, ESPCI Paris, LPC,CNRS,UMR 8249, 10 rue Vauquelin, F-75005 Paris, France
[2] Univ Paris Cite, Inst Pasteur, Spacial Regulat Genome Grp, CNRS 3525-25-28 Rue Dr Roux, F-75015 Paris, France
关键词
SACCHAROMYCES-CEREVISIAE; PCR; DATABASE; BAYANUS; VOLUME;
D O I
10.1039/d4lc00919c
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Combining different "omics" approaches, such as genomics and proteomics, is necessary to generate a detailed and complete insight into microbiome comprehension. Proper sample collection and processing and accurate analytical methods are crucial in generating reliable data. We previously developed the ChipFilter device for proteomic analysis of microbial samples. We have shown that this device coupled to LC-MS/MS can successfully be used to identify microbial proteins. In the present work, we have developed our workflow to analyze concomitantly proteins and nucleic acids from the same sample. We performed lysis and proteolysis in the device using cultures of E. coli, B. subtilis, and S. cerevisiae. After peptide recovery for LC-MS/MS analysis, DNA from the same samples was recovered and successfully amplified by PCR for the 3 species. This workflow was further extended to a complex microbial mixture of known compositions. Protein analysis was carried out, enabling the identification of more than 5000 proteins. The recovered DNA was sequenced, performing comparable to DNA extracted with a commercial kit without proteolysis. Our results show that the ChipFilter device is suited to prepare samples for parallel proteomic and genomic analyses, which is particularly relevant in the case of low-abundant samples and drastically reduces sampling bias.
引用
收藏
页码:590 / 599
页数:10
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