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A streamlined protocol for extracting RNA and genomic DNA from archived human blood and muscle
Authors:Gipsy Majumdar  Santiago Vera  Marshall B. Elam  Rajendra Raghow
Affiliation:1. Department of Veterans Affairs Medical Center, Memphis, TN 38104, USA;2. Department of Pharmacology, College of Medicine, University of Tennessee Health Science Center, Memphis, TN 38163, USA
Abstract:We combined the TRIzol method of nucleic acid extraction with QIAamp columns to achieve coextraction of RNA and genomic DNA from peripheral blood mononuclear cells (PBMCs) and biopsied skeletal muscle, both stored at −80 °C for many months. Total RNA was recovered from the upper aqueous phase of TRIzol. The interphase and organic phases were precipitated with ethanol, digested with proteinase K, and filtered through QIAamp MinElute columns to recover DNA. The combined protocol yielded excellent quality and quantity of nucleic acids from archived human PBMCs and muscle and may be easily adapted for other tissues.
Keywords:TRIzol   QIAamp   DNA/RNA extraction   Peripheral blood mononuclear cells   Skeletal muscle biopsy
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