Single-Cell RNA Barcoding and Sequencing

SCRB-Seq is a cost-efficient, multiplexed, single-cell mRNA sequencing technique (Soumillon et al., 2014). SCRB-Seq isolates single cells into wells using FACS. After cell lysis, poly(A)+ mRNAs are annealed to a custom primer containing a poly(T) tract, UMI, well barcode, and biotin. Template-switching RT and PCR amplification reactions are carried out on the mRNA, generating barcoded, full-length cDNA. cDNA strands from all wells are pooled together to be purified. They are PCR-amplified and purified further. The cDNA libraries are prepared using the Nextera XT library preparation protocol, with modified i5 primers. The resultant cDNA fragments are size-selected for 300_800 bp and sequenced.


  • Cost-efficient, high-throughput, single-cell transcriptome profiling
  • Highly sensitive gene-detection results compared to popular scRNA-Seq techniques (Ziegenhain et al., 2016)



Illumina Library prep and Array Kit Selector


Ziegenhain C., Parekh S., Vieth B., Smets M., Leonhardt H., et al. Comparative analysis of single-cell RNA-sequencing methods. bioRxiv. 2016;

Shapiro E., Biezuner T. and Linnarsson S. Single-cell sequencing-based technologies will revolutionize whole-organism science. Nat Rev Genet. 2013;14:618-630


Cacchiarelli D., Trapnell C., Ziller M. J., et al. Integrative Analyses of Human Reprogramming Reveal Dynamic Nature of Induced Pluripotency. Cell. 2015;162:412-424