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.
机构:
Chinese Acad Agr Sci, Biotechnol Res Inst, Beijing 100081, Peoples R ChinaChinese Acad Agr Sci, Biotechnol Res Inst, Beijing 100081, Peoples R China
机构:
Sun Yat Sen Univ, Inst Resp Dis, Div Pulm & Crit Care Med, Affiliated Hosp 1, Guangzhou, Guangdong, Peoples R ChinaSun Yat Sen Univ, Inst Resp Dis, Div Pulm & Crit Care Med, Affiliated Hosp 1, Guangzhou, Guangdong, Peoples R China
Kasimu, Kaidirina
Cui, Wenwen
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机构:
Shenyang Med Coll, Sch Basic Med, Shenyang, Peoples R ChinaSun Yat Sen Univ, Inst Resp Dis, Div Pulm & Crit Care Med, Affiliated Hosp 1, Guangzhou, Guangdong, Peoples R China
Cui, Wenwen
Wang, Yihan
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机构:
Shenyang Med Coll, Sch Basic Med, Shenyang, Peoples R ChinaSun Yat Sen Univ, Inst Resp Dis, Div Pulm & Crit Care Med, Affiliated Hosp 1, Guangzhou, Guangdong, Peoples R China
Wang, Yihan
Li, Xin
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机构:
Shenyang Med Coll, Sch Stomatol, Shenyang, Peoples R ChinaSun Yat Sen Univ, Inst Resp Dis, Div Pulm & Crit Care Med, Affiliated Hosp 1, Guangzhou, Guangdong, Peoples R China
Li, Xin
Wang, Hongbo
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机构:
Shenyang Med Coll, Sch Basic Med, Dept Human Anat, Shenyang, Peoples R China
Key Lab Human Ethn Specif & Phen Crit Illness Liao, Shenyang, Peoples R China
Key Lab Phen Res Shenyang, Shenyang, Peoples R ChinaSun Yat Sen Univ, Inst Resp Dis, Div Pulm & Crit Care Med, Affiliated Hosp 1, Guangzhou, Guangdong, Peoples R China
Wang, Hongbo
Yu, Xiaotong
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机构:
Shenyang Med Coll, Sch Basic Med, Dept Human Anat, Shenyang, Peoples R China
Key Lab Human Ethn Specif & Phen Crit Illness Liao, Shenyang, Peoples R China
Key Lab Phen Res Shenyang, Shenyang, Peoples R ChinaSun Yat Sen Univ, Inst Resp Dis, Div Pulm & Crit Care Med, Affiliated Hosp 1, Guangzhou, Guangdong, Peoples R China
Yu, Xiaotong
Ren, Fu
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机构:
Shenyang Med Coll, Sch Basic Med, Dept Human Anat, Shenyang, Peoples R China
Key Lab Human Ethn Specif & Phen Crit Illness Liao, Shenyang, Peoples R China
Key Lab Phen Res Shenyang, Shenyang, Peoples R ChinaSun Yat Sen Univ, Inst Resp Dis, Div Pulm & Crit Care Med, Affiliated Hosp 1, Guangzhou, Guangdong, Peoples R China
Ren, Fu
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