The below diagram is a representation of a barcoded double-stranded cDNA molecule produced from the 10X Genomics 3′ Gene Expression assay. This 5-minute video shows how Illumina’s “sequencing by synthesis” approach works and may be …
Single Cell RNA-seq
Interpreting the “fraction of reads in cells” values reported with the MiSeq results in single cell RNA-seq
In short: the “fraction of reads in cells” value is a measure of the proportion of sequencing reads assigned to true cells versus reads attributed to “background” such as RNA from dying/lysed cells captured in …
What is “On-Chip Multiplexing” in 10X Genomics?
10X Genomics recently launched new versions of their flagship 3′ and 5′ Gene Expression assays integrating an approach they call “On-Chip Multiplexing” or OCM. The OCM assays feature a relatively very low per sample reagent …
Sample Multiplexing vs. Library Multiplexing in 10X Genomics Single Cell RNA-seq
The below diagram illustrates the flow of a set of cell suspension samples through the process of library preparation and sequencing. A cell suspension, which can either be its own unique sample or (optionally) a …
Sample Prep for Single Cell/Nuclei RNA-seq
With rare exception, the single most important factor in determining the success of a single cell RNA-seq library preparation is the condition and health of the single cell/nuclei suspension at the time their transcripts are …
For single cell RNA-seq, why do sequencing cost estimates include both 2×150 bp and 2×50 bp read length options?
Most of the deep sequencing done in the UWBC DNA Sequencing Facility will be done on “10B” flow cells for the NovaSeq X+ instrument. These flow cells comprise eight lanes, each providing an estimated 1.25B …