SeekOne® Digital Droplet (SeekOne® DD) High throughput Single Cell Full-length RNA Sequence Transcriptome-seq (scFAST-seq) Kit, selfdeveloped by Beijing SeekGene BioSciences Co., Ltd., is a powerful commercial tool for high-throughput whole transcriptome profiling. The scFAST-seq method makes use of innovative techniques including semi-random primers, efficient reverse transcription, template swapping, and effective rRNA removal to build full-length RNA libraries of up to 12,000 cells.
Compared to conventional 3' scRNA-seq, scFASTseq has distinct advantages in detecting nonpolyadenylated transcripts, transcript coverage length, and identification of more splice junctions. With target region enrichment, scFAST-seq can simultaneously detect somatic mutations and cell states in individual tumour cells, providing valuable information for precision medicine.
The kit is designed to be used in conjunction with our SeekOne® Digital Droplet System (SeekOne® DD) to complete the entire process from single-cell nucleic acid labelling to transcriptome library construction. When equipped with "SeekSoul Tools", our single-cell data analysis software, we provide a one-stop-shop single-cell transcriptome solution.
Single Cell Full-length RNA Sequence Transcriptome-seq (scFAST-seq) allows full-length transcriptome analysis of thousands of fresh or fixed cells in a single experiment. In contrast to the current single-cell transcriptome assays based on oligo-dT reverse transcription, the method used in scFAST-seq provides coverage of the whole transcriptome. The sensitivity of the scFAST-seq technique assays depends primarily on the efficiency of reverse transcription, cDNA enrichment, and ribosome depletion.
First, cells and barcoded beads are added and react to generate emulsion droplets in the shaped channel of the SeekOne® Digital Droplet System. Reverse transcription is then performed by encapsulating the microbeads coupled to cell labels and semi-random primers with individual cells in a single droplet, resulting in cell-labelled cDNA fragments. Once the reverse transcription of each cDNA is complete, a template transition sequence (template switch oligo, TSO) is added to the 3' end of the cDNA using the end-transferase activity of the reverse transcriptase. The barcoded cDNA is then amplified in vitro, where the ribosomal RNA (rRNA) is depleted. Finally, a portion of the cDNA is fragmented, end-repaired, and ligated to a sequencing adaptor. The library is constructed and amplified using the unique dual-indexed polymerase chain reaction (PCR) primers for sequencing on Illumina or MGI platforms. In this way, the cDNA amplified by PCR would reflect both polyA and nonpolyA RNA sequences.