Open AccessHigh-yield skeletal muscle protein recovery from TRIzol after RNA and DNA extraction
Author(s) -
Yan Wen,
Ivan J. Vechetti,
Taylor Valentino,
John J. McCarthy
Publication year - 2020
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2020-0083
Subject(s) - trizol , chromatography , lysis , homogenization (climate) , protein purification , rna , extraction (chemistry) , centrifugation , dna , cell disruption , rna extraction , yield (engineering) , chemistry , biology , microbiology and biotechnology , biochemistry , gene , materials science , biodiversity , ecology , metallurgy
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.