RT Journal Article
SR Electronic
T1 RNA-seq library preparation from single pancreatic acinar cells
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 085696
DO 10.1101/085696
A1 Damian Wollny
A1 Sheng Zhao
A1 Ana Martin-Villalba
YR 2016
UL http://biorxiv.org/content/early/2016/11/04/085696.abstract
AB Single cell RNA sequencing technology has emerged as a promising tool to uncover previously neglected cellular heterogeneity. Multiple methods and protocols have been developed to apply single cell sequencing to different cell types from various organs. However, library preparation for RNA sequencing remains challenging for cell types with high RNAse content due to rapid degradation of endogenous RNA molecules upon cell lysis. To this end, we developed a protocol based on the SMART-seq2 technology for single cell RNA sequencing of pancreatic acinar cells, the cell type with one of the highest ribonuclease concentration measured to date. This protocol reliably produces high quality libraries from single acinar cells reaching a total of 5x106 reads / cell and ∼ 80% transcript mapping rate with no detectable 3´end bias. Thus, our protocol makes single cell transcriptomics accessible to cell type with very high RNAse content.