Low expression of key transcripts in scRNAseq dataset after deeper sequencing

Low expression of key transcripts in scRNAseq dataset after deeper sequencing

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I am having an issue with my single cell data set. This alert was shown after I requested additional reads per cell to be incorporated with the initial data we received. 10X output

We have previously performed single cell on this tissue so I am pretty certain that the issue is not due to trying to identify rare transcripts. We sent this data for sequencing to further test a hypothesis that a specific cell population is a major “expresser” of a gene of interest. We have shown this in multiple ways so far. I am seeing trends in the data that is in line with what we have previously found but the cell count is so low that my violin plots do not give a violin and I have sparse dots on the umap. See below. the goal is to show that genes 1,2 and 3 are expressed in the same cluster, cluster 0. Gene 3 is a cell-type identifier

the goal is to show that genes 1,2 and 3 are expressed in the same cluster, cluster 0. I am at my wits end with trying to represent this data. We have tried doing deeper sequencing to no avail. Does anyone have any idea is there is a way to represent this data as is in a way that makes sense. I don’t think violin plots or umap plots will be useful here. Like I said, this is not our only piece of data so we are pretty confident that the finding is real.

Also, on the sequencing/10X side, is there anyway to improve data quality given this type of error? Note that we did not observe this error (picture 1) on the first round of sequencing.


genomics


violin


single-cell


10X


plot

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