RNA capture pin technology: investigating long-term stability and mRNA purification specificity of oligonucleotide immobilization on gold and streptavidin surfaces

Advancing biomedical studies necessitates the development of cutting-edge technologies for the rapid extraction of nucleic acid. We characterized an RNA capture pin (RCP) tool that is non-destructive to the sample and enables rapid purification and enrichment of mRNA for subsequent genetic analysis. At the core of this technology is a pin (200 & mu;m x 3 cm) functionalized with dT15 capture sequences that hybridize to mRNA within 2 min of insertion in the specimen. Two methods for immobilizing the oligos on the surface of the RCPs were investigated: gold-thiol and biotin-streptavidin. The RNA capture efficiency of the RCPs was assessed using a radish plant. The average reverse transcription-quantitative polymerase chain reaction (RT-qPCR) cycle amplification values were 19.93 and 24.84 for gold- and streptavidin-coated pins, respectively. The amount of RNA present on the surface of the probes was measured using the Agilent 2100 Bioanalyzer. RNA sequencing was performed to determine the mRNA selectivity of the RNA capture pin. Gene read count analysis confirmed that the RNA purified via the gold-plated RCPs contained 70% messenger RNA, 10% ribosomal RNA, and 20% non-coding RNA. The long-term stability of the bond between the dT15 oligos and the surface of the RCPs was assessed over 4 months. A significant decrease in the dT15 surface coverage of the streptavidin-coated RCPs was observed after 2 weeks of storage at 4 & DEG;C. The gold-thiol RNA capture pins exhibited a retention rate of 40% of the oligos after 4 months of storage.

Automated summary

Advancing biomedical studies requires the development of advanced technologies for the rapid extraction of nucleic acid. We characterized an RNA capture pin (RCP) tool that enables non-destructive, rapid purification and enrichment of mRNA for genetic analysis. The RCP, functionalized with dT15 capture sequences, demonstrated high RNA capture efficiency and selectivity, with 70% messenger RNA, 10% ribosomal RNA, and 20% non-coding RNA. Evaluation of long-term stability showed that gold-thiol RNA capture pins retained 40% of the oligos after 4 months of storage, while streptavidin-coated pins showed a significant decrease in dT15 surface coverage after 2 weeks of storage at 4°C.