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Methods for simultaneous and quantitative isolation of mitochondrial DNA, nuclear DNA and RNA from mammalian cells

J. Nakhle, T. Özkan, K. Lněničková, P. Briolotti, ML. Vignais

. 2020 ; 69 (6) : 436-442. [pub] 20201026

Jazyk angličtina Země Velká Británie

Typ dokumentu časopisecké články, práce podpořená grantem

Perzistentní odkaz   https://www.medvik.cz/link/bmc22004765

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

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.

Citace poskytuje Crossref.org

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