Back

High throughput detection of variation in single-cell whole transcriptome through streamlined scFAST-seq

Jiao, S.; Zhao, Y.; Gong, Y.; Sang, G.; Chen, J.; Zhao, M.; Shi, H.; Han, J.; Sun, J.; Guan, Y.; Ma, X.; Zhang, G.

2023-03-23 genomics
10.1101/2023.03.19.533382 bioRxiv
Show abstract

High-throughput single-cell full-length RNA sequencing is a powerful tool to explore the entire transcriptome, including non-polyadenylated transcripts. We developed a single cell Full-length RNA Sequence Transcriptome sequencing method (scFAST-seq), which combines semi-random primers with high reverse transcription efficiency, template switching and convenient rRNA removal methods, allowing the construction of full-length RNA libraries of up to 12,000 cells within 8 hours. Compared to regular 3 scRNA-seq, scFAST-seq has similar sensitivity to mRNA detection, sequencing cost and experimental workflow. Moreover, scFAST-seq has clear advantages in detecting non-polyadenylated transcripts, covering longer transcript length, and identifying more splice junctions. In addition, scFAST-seq can more accurately predict the direction of cell differentiation by calculating RNA velocity. Furthermore, we demonstrated that scFAST-seq combined with target region enrichment can simultaneously identify somatic mutations and cellular status of individual tumor cells, which is valuable information for precision medicine.

Matching journals

The top 8 journals account for 50% of the predicted probability mass.

50% of probability mass above