A streamlined protocol for extracting RNA and genomic DNA from archived human blood and muscle
| Autor: | Marshall B. Elam, Santiago R. Vera, Rajendra Raghow, Gipsy Majumdar |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
Biophysics
Tissue Banks Biology Biochemistry Peripheral blood mononuclear cell chemistry.chemical_compound medicine Humans Molecular Biology Human blood Genome Human Muscles RNA Skeletal muscle DNA Cell Biology Molecular biology genomic DNA medicine.anatomical_structure chemistry Trizol Leukocytes Mononuclear Nucleic acid |
| Zdroj: | Analytical Biochemistry. 474:25-27 |
| ISSN: | 0003-2697 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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