A Split Bio kit allows for profiling of 100,000 cells among up to 48 distinct cell samples. After fixation, cells or nuclei undergo four rounds of combinatorial indexing. Cells are first distributed in 48 wells where reverse transcription produces cDNA with well-specific barcodes. Two subsequent rounds of pooling and splitting into 96 well plates ligate two additional cDNA barcodes. Finally, cell lysis and PCR amplification in eight distinct sublibraries adds a fourth and final barcode. Together, this yields a total of 48x96x96x8 possible barcode combinations, or the potential to uniquely profile 3,538,944 single cells. With additional rounds of split-pooling and an increase in the number of barcodes in each round, our method has the potential to scale to profile tens of millions of cells. The scalability of our technology is limited only by the diminishing costs of next generation sequencing.
With Split Bio, you will design your own single cell RNA-seq experiment that will cater to your individual conditions and timeline. You can assess up to 48 unique samples in parallel with our kit, giving you the space to test varying conditions or time points efficiently. Samples can be frozen and stored after cell fixation, and thus can be prepared separately and processed together. Additionally, before cell lysis you will divide your experiment into up to 8 sublibraries that will contain your choice of cell number, and they can be sequenced individually. The use of sublibraries allows for the freedom to adjust cell counts and sequencing depth. The flexibility of our kit improves time management, limits the need for repeat experiments, and expands the diversity and efficiency of scRNA-seq.
Other single cell RNA sequencing methods require complex equipment to partition individual cells and barcode RNA molecules in a cell-specific manner. Our Single Cell Whole Transcriptome Kit can outperform these methods while requiring only basic laboratory tools. By using cells themselves as compartments, we can employ a series of split-pooling steps to label all RNA molecules in a given cell with a unique combination of barcodes. We provide all the required materials in our affordable kit, and thus we limit the need for instrument maintenance or troubleshooting and ensure reliable results for every experiment.