Getting started with Cell Ranger

Cell Ranger is a set of analysis pipelines that process Chromium single cell 3′ RNA-seq data. The pipelines process raw sequencing output, performs read alignment, generate gene-cell matrices, and can perform downstream analyses such as clustering and gene expression analysis. Cell Ranger includes four pipelines: cellranger mkfastq cellranger count cellranger aggr cellranger reanalyze You can…

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10x single cell BAM files

The Chromium Single Cell 3′ Solution is a commercial platform developed by 10x Genomics for preparing single cell cDNA libraries for performing single cell RNA-seq. In addition, 10x Genomics have developed an entire software suite called Cell Ranger that can process the raw BCL files produced by an Illumina sequencer and output a final gene-barcode…

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Merging two 10x single cell datasets

I was going to write a post on using the Seurat alignment method as a batch correction tool but as it turned out the two datasets that I chose didn’t seem to have strong batch effects! I heard about the alignment method sometime last year but was motivated to try it out after listening to…

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Getting started with Monocle

Monocle is an R package developed for analysing single cell gene expression data. Specifically, the package provides functionality for clustering and classifying single cells, conducting differential expression analyses, and constructing and investigating inferred developmental trajectories. The toolkit provides various alternative approaches for each analysis, hence your workflow may differ from the approach I’ve taken in…

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Getting started with Seurat

Updated 2018 April 11th This post follows the Peripheral Blood Mononuclear Cells (PBMCs) tutorial for 2,700 single cells. It was written while I was going through the tutorial and contains my notes. The dataset for this tutorial can be downloaded from the 10X Genomics dataset page but it is also hosted on Amazon (see below)….

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