Comments (4)
I think UMI_tools
has methods for figuring out "true" barcodes from mismatches that could be applied to the viral barcodes within single cells.
Within the whole mix, there is some complicated set of viral barcodes. That is true both if we look at all barcodes over transcriptomics (across cells) or if we look at all barcodes in the supernatant.
We really only care about the viral barcodes that are in the transcriptomics.
Furthermore, within each cell, we expect there to be only at most a few distinct viral barcodes.
My suggestion is that first we identify truly infected cells in the transcriptomics.
Then we use the transcriptomic viral barcode calls to use something like UMI_tools to figure out what are the true viral barcode in that cell. And then we have our set of true viral barcodes in the transcriptomic data.
from barcoded_flu_pdmh1n1.
If we take this approach, we need to #58
from barcoded_flu_pdmh1n1.
A few ideas came out of our discussion about this issue today:
- As you noted above, in a low MOI infection, each cell should only have one or a few distinct viral barcodes. We can use UMI tools to correct all the barcodes within a cell in the transcriptomics data.
- I expect the edit distance between true barcodes to be quite large. We are only sampling a few thousand barcodes from a potential nt space of billions. I will try to plot the edit distance between each pairwise comparison.
- I will examine the progeny viral barcode data to see if barcodes are ever shared between WT and dblSyn variant libraries. The infections and sequencing for the two viral tags are handled totally independently. If they are fully unique, we can use the barcode information to assign tag identities to cells.
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This nebulous issue has been superseded by more specific tasks. I am closing it.
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Related Issues (20)
- Sequence of unusual viral barcodes HOT 1
- Number of transcript counts per viral barcode HOT 1
- Viral barcode rank within cell
- STARsolo version HOT 1
- Total number of transcripts per cell and effect on viral barcodes HOT 1
- update `STAR`?
- Annotate viral barcodes that infect at least 1 cell
- Annotate cells with complete genomes/missing genes
- Plot viral burden vs viral barcode abundance
- Calculate viral barcode saturation in transcriptome HOT 1
- UMIs per viral barcode in trascriptome HOT 1
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- error with `Snakefile`
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- viral mutations with negative numbers? HOT 2
- Put flu genes in canonical order
- Report file missing: Assign infection Status HOT 4
- High MOI interactive plot is not displayed HOT 3
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