Accurate quantification of RNA concentration

Protocol Overview

1. Make working solution of dye. Mix 199 ul buffer with 1 ul of dye to make 200 times dilution. If there are x samples, plus 2 standards, there will be (x+2) samples in total. Then multiply 199 and 1 with (x+2). For examples, if 2 samples, then mix 199×4 ul buffer and 4 ul dye together by pipetting several times. 2. For samples, mix 199 ul of working solution with 1 ul of RNA samples (200 times dilution). Then vortex very gently for 3 s and incubate at room temperature for 2 mins. 3. For standard, mix 190 ul of working solution with 10 ul of each standard (20 times dilution). Then vortex very gently for 3 s and incubate at room temperature for 2 mins.4. Activate Qubit -> RNA -> BR (Broad range) RNA -> New standard “Yes” -> insert standard 1 (0) -> Read -> insert standard 2 (100 ng/ul) -> Read -> insert samples -> Read -> calculate stock concentration, scroll to select the added volume of RNA sample.