The IsoBind VineVantage RNA Kit provides viticulture, plant-health and research laboratories with a manual silica spin-column workflow for total RNA purification from frozen grapevine leaf and berry tissue.
Samples are lysed in 500 µL Nova RG with 10 µL BME at 56°C for 8 minutes before centrifugation and filter-column clarification. Optional PolyTrap treatment may be incorporated during lysis where additional matrix clean-up is required. Prime R2 is added at 0.3 times the recovered lysate volume, followed by 20 µL Booster L and Bind C at 0.2 times the lysate volume. A 15-minute freezer precipitation stage prepares the clarified lysate for binding to the blue silica spin column.
One 600 µL Wash Nova-Y stage and two 600 µL Wash Ultimus stages reduce contaminants before the column is dried. Optional on-column DNase I treatment may be incorporated when additional genomic DNA removal is required, and DNase I is included with the kit. RNA is recovered in 50 µL Elution Buffer preheated to approximately 60°C.
The workflow processes 80 to 100 mg frozen grapevine leaf or berry tissue in approximately 45 minutes using a standard laboratory centrifuge. Petiole tissue requires an additional chloroform clean-up and should be treated as a qualified secondary sample. Cane and other woody vine tissues are not currently supported.
Purified RNA is suitable for RT-qPCR, gene expression studies, transcriptomics and next generation sequencing. Responsive technical support is available for grapevine sample preparation and workflow setup.
