Methods for simultaneous and quantitative isolation of mitochondrial DNA, nuclear DNA and RNA from mammalian cells
Jazyk angličtina Země Velká Británie, Anglie Médium print-electronic
Typ dokumentu časopisecké články, práce podpořená grantem
Grantová podpora
French Ministry of Research (MESRI)
Ames Research Center NASA - United States
Scientific and Technological Research Council of Turkey (TUBITAK)
Ligue Contre le Cancer-Comité du Gard
CZ.02.2.69/0.0/0.0/16_027/0008482
Mobility Support project at UP
PubMed
33103926
DOI
10.2144/btn-2020-0114
Knihovny.cz E-zdroje
- Klíčová slova
- RNA, mammalian cells, mitochondria, mitochondrial DNA (mtDNA), nuclear DNA (ncDNA),
- MeSH
- biochemie metody MeSH
- buněčné jádro metabolismus MeSH
- buňky Hep G2 MeSH
- glioblastom metabolismus patologie MeSH
- lidé MeSH
- messenger RNA izolace a purifikace MeSH
- mitochondriální DNA izolace a purifikace MeSH
- nádorové kmenové buňky metabolismus MeSH
- reagenční diagnostické soupravy MeSH
- RNA izolace a purifikace MeSH
- savci metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- messenger RNA MeSH
- mitochondriální DNA MeSH
- reagenční diagnostické soupravy MeSH
- RNA MeSH
The aim of this study was to assess two protocols for their capacities to simultaneously isolate RNA, mtDNA and ncDNA from mammalian cells. We compared the Invitrogen TRIzol-based method and Qiagen DNeasy columns, using the HepG2 cell line and human primary glioblastoma stem cells. Both methods allowed the isolation of all three types of nucleic acids and provided similar yields in mtDNA. However, the yield in ncDNA was more than tenfold higher on columns, as observed for both cell types. Conversely, the TRIzol method proved more reproducible and was the method of choice for isolating RNA from glioblastoma cells, as demonstrated for the housekeeping genes RPLP0 and RPS9.
Faculty of Medicine Department of Medical Biology University of Ankara Ankara Turkey
Institute of Molecular Genetics of Montpellier CNRS Univ Montpellier F 34090 Montpellier France
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