Phenol/Chloroform-Free TiO 2 -Based miRNA Extraction from Cell Lysate.
Denisa SmelaEliska ZelinkovaPavel RehulkaZuzana BilkovaRudolf KupcikPublished in: International journal of molecular sciences (2022)
While microRNAs are considered as excellent biomarkers of various diseases, there are still several remaining challenges regarding their isolation. In this study, we aimed to design a novel RNA isolation method that would help to overcome those challenges. Therefore, we present a novel phenol/chloroform-free, low-cost method for miRNA extraction. Within this method, RNA is extracted from cell lysate with an isopropanol/water/NaCl system, followed by solid-phase extraction using TiO 2 microspheres to effectively separate short RNAs from long RNA molecules. We also demonstrated the pH-dependent selectivity of TiO 2 microspheres towards different sizes of RNA. We were able to regulate the size range of extracted RNAs with simple adjustments in binding conditions used during the solid-phase extraction.
Keyphrases
- solid phase extraction
- molecularly imprinted
- liquid chromatography tandem mass spectrometry
- high performance liquid chromatography
- low cost
- tandem mass spectrometry
- simultaneous determination
- liquid chromatography
- single cell
- cell therapy
- gas chromatography
- quantum dots
- ultra high performance liquid chromatography
- nucleic acid
- mass spectrometry
- visible light
- high resolution
- ms ms
- transcription factor