279002 Quantification of Nucleic Acid Hybridization near Solid Liquid Interface
Given low concentration of target RNA in biological samples, RNA isolation methods needs fundamental understanding of hybridization process near interfaces. In this study, we present a novel microfludic method for sDNA isolation near solid liquid interface. We have examined the key factors that have effects on the hybridization efficiency, which include incubation time (15min, 25min, and 35min), temperature (25C, 40C and 70C) and location of target sequence (5’ end, middle, and 3’ end) in sDNA. We present extensive results on the capacity of solid liquid interfaces in capturing RNA targets that incorporates the advantages of microfluidics.
We found that there is minimal incubation time for optimal hybridization to happen and beyond that there is no significant change. We also found that heating the solid liquid system will enhance the hybridization efficiency. There is 3-fold increase in hybridization capacity with 40C treatment and 2-fold increase with 70C treatment after 15min incubation compares to that in room temperature. This study is useful for examining target sDNA hybridization capacity with biotinylated capture near solid liquid surface. This investigation presents the parameters of this quick method on a microchip platform with many possible applications. The technique can be used to improve a number of assays, including DNA/RNA isolation and amplification, blood plasma separation and peptide isolation. Our study provides a quantitative guidance to clinically relevant RNA detection methods.
Acknowledgements: We acknowledge support from the National Institute of Health (US) and the China Scholarship Council.