What is the recommended volume of Reaction Buffer for various amounts of cells when using SMARTer Ultra Low kits?
What is the recommended volume of Reaction Buffer for various amounts of cells when using SMARTer Ultra Low kits?
For cell lysis using the SMART-Seq mRNA Kit for Sequencing, we recommend the addition of 1 µl of 10X Reaction Buffer followed by a 5-minute incubation at room temperature. For the legacy kits, the amount of Reaction Buffer will vary depending on the input volume of your RNA or cell sample. Maintaining the same volume of Reaction Buffer for all cell samples is not necessary. Do not add more than 5 µl of Reaction Buffer to your sample. If necessary, add nuclease-free water to a final volume of 10 µl. Allow lysis to proceed for 5 minutes at room temperature or on ice (4°C).