Isolation and Solubilization of Proteins After TRI zol ® Extraction of RNA and DNA from Patient Material Following Prolonged Storage
Author(s) -
Amanda B. Hummon,
Sharlene Lim,
Michael J. Difilippantonio,
Thomas Ried
Publication year - 2007
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000112401
Subject(s) - trizol , proteome , rna , rna extraction , dna , genomic dna , gel electrophoresis , polyacrylamide gel electrophoresis , dna extraction , microbiology and biotechnology , western blot , biology , solubilization , chemistry , chromatography , biochemistry , gene , polymerase chain reaction , enzyme
A systems approach is being applied in many areas of the biological sciences, particularly in cancer research. The coordinated, simultaneous extraction of DNA, RNA, and proteins from a single sample is crucial for accurate correlations between genomic aberrations and their consequences on the transcriptome and proteome. We present an approach to extract and completely solubilize up to 98% of the total protein recovered from archived samples following TRIzoL isolation of RNA and DNA. We also demonstrate using polyacrylamide gel electrophoresis (PAGE) and Western blot analysis that the proteins, representing both a wide molecular weight range and some posttranslational modifications, such as protein phosphorylation, remain stable in phenol-ethanol for up to 3 years at -20 degrees C.
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