Single-nuclei RNA sequencing characterizes cell types at the gene level. However, compared to single-cell approaches, many single-nuclei cDNAs are purely intronic, lack barcodes and hinder the study of isoforms. Here we present single-nuclei isoform RNA sequencing (SnISOr-Seq). Using microfluidics...
Single-nuclei RNA sequencing characterizes cell types at the gene level. However, compared to single-cell approaches, many single-nuclei cDNAs are purely intronic, lack barcodes and hinder the study of isoforms. Here we present single-nuclei isoform RNA sequencing (SnISOr-Seq). Using microfluidics...
which grouped all nuclei into 6 non-neuronal and 23 neuronal (12 glutamatergic, 1 cholinergic, 10 GABAergic) major cell clusters (Fig.1b), using well-established marker genes (Fig.1c–e). We identified three neuroglia clusters, specifically one immature and one myelinating/mature oligodendrocyt...
Single-cell sequencing provides analysis of thousands of individual cells within complex environments, as opposed to traditional bulk sequencing methods that only provide a representative average of all cells. With developments in microfluidics and molecular barcoding, NGS-based technologies have increasingly ...
this technology is constrained to the analyses of fresh tissue. Single-nuclei RNA sequencing (snuclRNA-seq) is still in its initial developmental stages. In this study, we showed that it is feasible to employ it to ascertain highly informative and unique brain tissue collected during many years...
(<half the total entropy possible) were later identified as either having few nuclei or being enriched for carriers of genetic factors (i.e., ADAD,MS4A, orTREM2). Neuronal cell states were then categorized as excitatory (EN) or inhibitory (IN; See Methods). We capture an average of ...
1 mouse brain using the Chromium Nuclei Isolation Kit and enriched to remove debris. Nuclei were loaded separately onto a Next GEM Chip G (5,000 nuclei) and a GEM-X 3' chip (5,000 nuclei) and were processed on the Chromium X followed by library preparation, sequencing, and data ...
nuclei,suchasenrichmentofcertaintranscriptsandprecursorsofsomenoncodingRNAs.By followingthisprocedure,ittakesabout4dtoconstructcDNAlibrariesthatarereadyfor sequencing. Reprintsandpermissionsinformationisavailableonlineathttp://.nature/reprints/index.html. CorrespondenceshouldbeaddressedtoR.S.L.(rlasken@jcvi)orR.V....
Single nuclei suspensions were barcoded using the 10x Chromium Single Cell platform, cDNA libraries generated and sequenced on Illumina platform and integrated together using RCPA. To build the heatmap we calculated the average mean expression of gene of interest within each cell type and across ...
We develop a method to isolate three nuclei from a pollen grain and four microspores from a tetrad for whole-genome sequencing. A high rate of aneuploidy is observed at the three-nucleus stage (6/22 pollens) rather than at the tetrad stage (1/72 microspores) in one HI line CAU5. ...