DESeq2モデル

RNA-Seq with Bioconductor in R

Mary Piper

Bioinformatics Consultant and Trainer

DESeq2モデル

DE理論

RNA-Seq with Bioconductor in R

DESeq2モデル - 平均と分散の関係

# Syntax for apply()
apply(data, rows/columns, function_to_apply)
# Calculating mean for each gene (each row)
mean_counts <- apply(wt_rawcounts[, 1:3], 1, mean)
# Calculating variance for each gene (each row)
variance_counts <- apply(wt_rawcounts[, 1:3], 1, var)
RNA-Seq with Bioconductor in R

DESeq2モデル - 分散

平均と分散の関係をプロット:

# Creating data frame with mean and variance for every gene
df <- data.frame(mean_counts, variance_counts)
ggplot(df) +
        geom_point(aes(x=mean_counts, y=variance_counts)) + 
        scale_y_log10() +
        scale_x_log10() +
        xlab("Mean counts per gene") +
        ylab("Variance per gene")
RNA-Seq with Bioconductor in R

DESeq2モデル - 分散

平均と分散の関係

RNA-Seq with Bioconductor in R

DESeq2モデル - 分散

$Var$: 分散

$\mu$: 平均

$\alpha$: 分散パラメータ

分散の式: $Var = \mu + \alpha * \mu^{2}$

平均・分散・分散パラメータの関係:

$$\uparrow variance \Rightarrow \uparrow dispersion$$

$$\uparrow mean \Rightarrow \downarrow dispersion$$

RNA-Seq with Bioconductor in R

DESeq2モデル - 分散

# Plot dispersion estimates
plotDispEsts(dds_wt)

分散プロット

RNA-Seq with Bioconductor in R

DESeq2モデル - 分散

不適切な分散プロット

RNA-Seq with Bioconductor in R

練習しましょう!

RNA-Seq with Bioconductor in R

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