Pool-and-Split Vessel and Synthesizer for the Synthesis of Barcoded Beads used in Single-Cell Applications

Pool and Split Vessel and Synthesizer for the Synthesis of Barcoded Beads used in Single Cell Applications Abstract SummaryAs the single cell genomics and in particular RNA sequencingscRNA Seqfield has maturedmethods to work in high throughput have been developedMany of these critically rely on solid support beads to capture cellular RNA targets in microwells or reverse emulsionsAn essential requirement on these beads is a DNA barcode that is approximately unique to each bead and ties the contents of that bead back to a single original cellThe DNA barcode synthesis requirements are not compatible with state of the art DNA synthesizersand as suchbarcode truncation and lack of complexity aboundimpacting method fidelityCurrent synthesized barcoded primer beads have quality issues that can be tracedin partto the manual handling of the beads during barcode generationorpool and splitstepsand to the incorporation inefficiency of nucleotides during the chemical synthesis of the growing oligonucleotide chainsWe propose to develop an automated pool and split system to synthesize uniquely barcoded primer beads with a high content of full length sequences by eliminating all manual handling and increasing overall bead coupling efficiency and recovery yieldsAbstract Project Summary docx PageofPool and Split Vessel and Synthesizer for the Synthesis of Barcoded Beads used in Single Cell Applications NarrativeWe will construct a pool and split reaction vessel and synthesizer that can automatically pool andsplit beads to generate higher quality single or multi function oligo barcoded beadsin a more cost and reagent effective manner while significantly reducing turn around timeImproving quality and reproducibility will accelerate the development and maturation of single cell technologies while ensuring that high quality profiling of large cell population at single cell resolution is not hindered by poorly synthesized cell barcodesNarrative docx Pageof