Validation of RNA extraction procedures focused on micro RNA expression analysis
Language English Country Czech Republic Media print
Document type Journal Article, Research Support, Non-U.S. Gov't, Validation Study
PubMed
23537528
PII: file/5671/FB2013A0006.pdf
Knihovny.cz E-resources
- MeSH
- Humans MeSH
- MicroRNAs genetics metabolism MeSH
- Molecular Biology methods MeSH
- Gene Expression Regulation MeSH
- Reproducibility of Results MeSH
- Gene Expression Profiling * MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Validation Study MeSH
- Names of Substances
- MicroRNAs MeSH
The sampling procedure is a crucial step in every kind of experiment. This is also true in gene expression profiling experiments, where high quality and sufficient quantity of extracted RNA plays a significant role in the experimental outcome. We have compared five different RNA extraction protocols from peripheral blood/PBMCs with the aim to define the most suitable method for the miRNA expression profiling experiments. Convincing results in terms of highest quantity and quality were obtained by the TRIzol-chloroform extraction method. The total RNA obtained using this method contained the highest portion of good-quality miRNA molecules, which was also confirmed by gene-specific real-time PCR experiments.
