Technical information describing isolation and absolute quantification of miRNAs from various biological matrices using BioVendor RNA Isolation kits and miREIA quantification kits.
Introduction
MicroRNAs (miRNAs) are small non-coding RNA molecules that regulate gene translation through silencing of target mRNAs. They are involved virtually in all biological processes. Numerous studies identified miRNAs as promising diagnostic and prognostic biomarkers of many human pathologies.
Recently, BioVendor has launched a novel method for absolute quantification of miRNA – microRNA Enzyme Immunoassay (miREIA). The goal of the present study was to examine performance of miREIA in diverse biological samples.
To this end, BioVendor miREIA kits were used for quantification of miRNAs isolated from whole blood, HeLa cells, peripheral blood mononuclear cells, colorectal cancer tissue, synovial fluid, saliva and urine. The results showed that BioVendor miREIA kits were suitable for quantification of miRNA in all the matrices mentioned above.
RESULTS
Whole blood
Whole blood was collected into PAXgene blood RNA collecting tubes. RNA was isolated using BioVendor RNA Isolation Kit for RNA isolation (RIK001), a kit intended for the isolation of RNA from cultured cells, human/animal tissues, plant tissues and whole blood. Absolute concentration of twelve different miRNAs was measured by BioVendor miREIA kits.
HeLa cells
RNA from HeLa cells was isolated using the BioVendor RNA Isolation Kit RIK001. Absolute concentration of eight different miRNAs was measured by BioVendor miREIA kits.
Peripheral blood mononuclear cells
Peripheral blood mononuclear cells (PBMC) were isolated from whole blood by density gradient centrifugation method using Histopaque. RNA was isolated using the BioVendor RNA Isolation Kit RIK001. Absolute concentration of seven different miRNAs was measured by BioVendor miREIA kits.
Colorectal cancer tissue
Colorectal cancer (CRC) tissue obtained by resection was stabilized and stored in RNAlater solution. RNA was isolated using the BioVendor RNA Isolation Kit RIK001. Absolute concentration of eight different miRNAs was measured by BioVendor miREIA kits.
Synovial fluid
RNA from synovial fluid was isolated using the BioVendor RNA Isolation Kit Plasma/Serum (RIK002). Absolute concentration of five different miRNAs was measured by BioVendor miREIA kits.
Saliva
RNA from saliva was isolated using the BioVendor RNA Isolation Kit RIK002. Absolute concentration of two different miRNAs was measured by BioVendor miREIA kits.
Urine
RNA from urine was isolated using the BioVendor RNA Isolation RIK002. Absolute concentration of two different miRNAs was measured by BioVendor miREIA kits.
CONCLUSIONS
Our pilot mini-study proved that miRNAs can be isolated from a wide range of biological matrices using BioVendor RNA Isolation Kits (RIK001 and RIK002). Subsequently selected miRNAs can be quantified using single miRNA-specific BioVendor miREIA kits. The tested matrices were: whole blood, HeLa cells, peripheral blood mononuclear cells (PBMC), colorectal cancer (CRC) tissue, synovial fluid, saliva, and urine.