High-yield skeletal muscle protein recovery from TRIzol after RNA and DNA extraction

Biotechniques. 2020 Oct;69(4):264-269. doi: 10.2144/btn-2020-0083. Epub 2020 Aug 11.

Abstract

Extraction of DNA, RNA and protein from the same sample would allow for direct comparison of genomic, transcriptomic and proteomic information. Commercially available kits exhibit poor protein yield and the TRIzol® reagent produces a protein pellet that is extremely difficult to solubilize. In response to these limitations, this study presents an optimized method for the extraction of protein from the organic phase of TRIzol that allows for higher yield recovery of skeletal muscle protein compared with direct homogenization in a common protein lysis buffer. The presented method is inexpensive, simple and fast, requires no additional treatment of the protein pellet for dissolution, and is compatible with downstream western blot applications.

Keywords: ethanol–bromochloropropane–water method; guanidinium thiocyanate–phenol–chloroform extraction; modified TRIzol protein isolation; skeletal muscle.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Blotting, Western
  • DNA / chemistry
  • DNA / isolation & purification*
  • Genomics
  • Guanidines / pharmacology*
  • Humans
  • Muscle Proteins / chemistry
  • Muscle Proteins / genetics
  • Muscle Proteins / isolation & purification*
  • Muscle, Skeletal / chemistry
  • Muscle, Skeletal / metabolism
  • Phenols / pharmacology*
  • RNA / chemistry
  • RNA / isolation & purification*

Substances

  • Guanidines
  • Muscle Proteins
  • Phenols
  • trizol
  • RNA
  • DNA