Comparison and optimization of methods for the simultaneous extraction of DNA, RNA, proteins, and metabolites

被引:19
作者
Vorreiter, Fraenze [1 ]
Richter, Silke [1 ]
Peter, Michel [1 ]
Baumann, Sven [1 ,2 ]
von Bergen, Martin [1 ,2 ,3 ]
Tomm, Janina M. [1 ]
机构
[1] UFZ Helmholtz Ctr Environm Res, Dept Mol Syst Biol, D-04318 Leipzig, Germany
[2] Univ Leipzig, Fac Biosci Pharm & Psychol, D-04103 Leipzig, Germany
[3] Aalborg Univ, Dept Chem & Biosci, DK-9220 Aalborg, Denmark
关键词
Molecular systems biology; Genomics; Transcriptomics; Proteomics; Metabolomics; SYSTEMS; CELLS;
D O I
10.1016/j.ab.2016.05.011
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The challenge of performing a time-resolved comprehensive analysis of molecular systems has led to the quest to optimize extraction methods. When the size of a biological sample is limited, there is demand for the simultaneous extraction of molecules representing the four areas of "omics": genomics, transcriptomics, proteomics, and metabolomics. Here we optimized a protocol for the simultaneous extraction of DNA, RNA, proteins, and metabolites and compared it with two existing protocols. Our optimization comprised the addition of a methanol/chloroform metabolite purification before the separation of DNA/RNA and proteins. Extracted DNA, RNA, proteins, and metabolites were quantitatively and/or qualitatively analyzed. Of the three methods, only the newly developed protocol yielded all biomolecule classes of adequate quantity and quality. (C) 2016 The Author(s). Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license.
引用
收藏
页码:25 / 33
页数:9
相关论文
共 21 条
[1]   Mutant T cell lines as model systems for the dissection of T cell antigen receptor signaling pathways [J].
Abraham, RT .
IMMUNOLOGIC RESEARCH, 2000, 22 (2-3) :95-117
[2]   Mass spectrometry-based proteomics [J].
Aebersold, R ;
Mann, M .
NATURE, 2003, 422 (6928) :198-207
[3]  
Baumann S., 2014, J INTEGR OMICS, V4, P11
[4]   Quantitative Evaluation of Intracellular Metabolite Extraction Techniques for Yeast Metabolomics [J].
Canelas, Andre B. ;
ten Pierick, Angela ;
Ras, Cor ;
Seifar, Reza M. ;
van Dam, Jan C. ;
van Gulik, Walter M. ;
Heijnen, Joseph J. .
ANALYTICAL CHEMISTRY, 2009, 81 (17) :7379-7389
[5]  
Fleige Simone, 2006, Molecular Aspects of Medicine, V27, P126, DOI 10.1016/j.mam.2005.12.003
[6]   Comparative analysis of the venom proteomes of Vipera ammodytes ammodytes and Vipera ammodytes meridionalis [J].
Georgieva, Dessislava ;
Risch, Michaela ;
Kardas, Anna ;
Buck, Friedrich ;
von Bergen, Martin ;
Betzel, Christian .
JOURNAL OF PROTEOME RESEARCH, 2008, 7 (03) :866-886
[7]  
Herbert Ben, 2009, V519, P47, DOI 10.1007/978-1-59745-281-6_4
[8]   A new approach to decoding life: Systems biology [J].
Ideker, T ;
Galitski, T ;
Hood, L .
ANNUAL REVIEW OF GENOMICS AND HUMAN GENETICS, 2001, 2 :343-372
[9]   The low molecular weight proteome of Halobacterium salinarum [J].
Klein, Christian ;
Aivaliotis, Michalis ;
Olsen, Jesper V. ;
Falb, Michaela ;
Besir, Hueseyin ;
Scheffer, Beatrix ;
Bisle, Birgit ;
Tebbe, Andreas ;
Konstantinidis, Kosta ;
Siedler, Frank ;
Pfeiffer, Friedhelm ;
Mann, Matthias ;
Oesterhelt, Dieter .
JOURNAL OF PROTEOME RESEARCH, 2007, 6 (04) :1510-1518
[10]   A SIMPLE METHOD FOR DISPLAYING THE HYDROPATHIC CHARACTER OF A PROTEIN [J].
KYTE, J ;
DOOLITTLE, RF .
JOURNAL OF MOLECULAR BIOLOGY, 1982, 157 (01) :105-132