Yes, aliquots of the same fixed sample can be used in multiple barcoding runs (Section 1).
If you are planning to use the same sample for multiple barcoding runs, at the end of fixation, cells should be split into several aliquots. Simply calculate the number of cells needed to input into the Round 1 plate, and aliquot the corresponding volume* of fixed cells into as many 1.5ml tubes (for volumes greater than 50 μl) or 0.5mL tubes (for volumes greater than 30 μl) as desired. Additionally, if possible, it is recommended to create a “counting aliquot” of approximately 10 μl that can be thawed in advance of barcoding for counting and quality control of the fixed and frozen sample. Once a fixed sample aliquot is thawed, it should not be refrozen.
Additional information on sample storage is available in the following article: How should I store my cells or nuclei after fixation?
*Note: The required concentration for fully loading a sample into the Round 1 plate varies depending on kit size. Cells should be resuspended at higher than the minimum concentration for the largest kit planned to be used, and aliquoted appropriately. Please refer to the EvercodeTM Fixation user guides for more information.