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

Anal Biochem. 2016 Sep 1:508:25-33. doi: 10.1016/j.ab.2016.05.011. Epub 2016 May 27.

Abstract

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.

Keywords: Genomics; Metabolomics; Molecular systems biology; Proteomics; Transcriptomics.

Publication types

  • Comparative Study

MeSH terms

  • Chemistry Techniques, Analytical / methods*
  • Computational Biology / methods*
  • DNA / chemistry
  • DNA / isolation & purification*
  • Proteins / chemistry
  • Proteins / isolation & purification*
  • Proteins / metabolism
  • RNA / chemistry
  • RNA / isolation & purification*

Substances

  • Proteins
  • RNA
  • DNA