For both cone- and starburst-enriched genes, gene expression values increased linearly with an increase in the of cones or starburst cells in the mixtures. The linear correlation coefficients were independent of the extent to which genes were expressed in the pure cone and starburst cell groups.
This suggests that A practical use of the finding that amplification is linear is to separate the transcriptome of a cell group that contains cells from different types into its cell-type components. Assuming, for example, a bi chat room group Group A containing a mixture of two cell types, Type 1 and Type 2, as well as a futher cell group Group B containing only Type 1 Supplementary Figure We wish to determine the transcriptome of Type 2 but we have no cell group Group C that contains Type 2 only.
Or more generally, assume a cell group Group A containing a mixture of cell types, Type 1, Type 2…Type N, as well as a further cell group Group B containing only Type 1. We wish to determine the mixed transcriptome of Group C that contains Type 2….
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Type N, without Type 1. This separation of cell-type transcriptomes is useful for two reasons. We wished to eliminate rod contamination.
Second, we found that cones in some mouse lines are labeled together with a type of amacrine cell and we needed to determine the amacrine cell transcriptome alone. Due to linearity, the following procedure can be used to separate Type 1 from the rest of the cell chaf in a mixture Group A.
A set of cell type-specific genes for Type 1 is determined. This may be known a priori or from the dataset itself.
The ratio of the expression value in Group A to the expression value in Group B is calculated for each Type 1-specific gene. The mean ratio is the estimate of the of Type 1 cells in Group A. Next, the transcriptome the expression values of all genes, which we treat as a vector of Group B is multiplied by the mean ratio and the result is subtracted from the transcriptome of Group A.
Note that if Group A contains only two cell types, Type 1 and Type 2, sexting tramping lake, saskatchewan masturbation the transcriptome of Group C is the transcriptome of Type 2 cells.
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We have demonstrated and quantitatively evaluated this procedure for the cone:starburst mixtures described above. Cones served as the Type 1 cells and starburst cells as Type 2.
In our experiment, we had one pure cone group and one pure starburst cell group as well as six mixtures, and First, we predicted the mixture composition using different s of cone-specific genes. Since we chose 20 cone-specific genes, the use of single genes presents 20 possible ways of predicting the of cones in the mixture.
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