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An optimized protocol for single cell transcriptional profiling by combinatorial indexing
- Publication Year :
- 2021
-
Abstract
- Single cell combinatorial indexing RNA sequencing (sci-RNA-seq) is a powerful method for recovering gene expression data from an exponentially scalable number of individual cells or nuclei. However, sci-RNA-seq is a complex protocol that has historically exhibited variable performance on different tissues, as well as lower sensitivity than alternative methods. Here we report a simplified, optimized version of the three-level sci-RNA-seq protocol that is faster, higher yield, more robust, and more sensitive, than the original sci-RNA-seq3 protocol, with reagent costs on the order of 1 cent per cell or less. We showcase the optimized protocol via whole organism analysis of an E16.5 mouse embryo, profiling ~380,000 nuclei in a single experiment. Finally, we introduce a "tiny sci-*" protocol for experiments where input is extremely limited.<br />fixed a couple errors
- Subjects :
- Genomics (q-bio.GN)
FOS: Biological sciences
Quantitative Biology - Genomics
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Accession number :
- edsair.doi.dedup.....b634df61e5cece6c172d84b826d7a43e