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andrewGhazi committed Sep 30, 2024
1 parent 2380fee commit a916d75
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Showing 5 changed files with 16 additions and 14 deletions.
2 changes: 1 addition & 1 deletion episodes/cell_type_annotation.Rmd
Original file line number Diff line number Diff line change
Expand Up @@ -650,7 +650,7 @@ Use `BiocParallel` and the `BPPARAM` argument! This example will set it to use f
library(BiocParallel)
my_bpparam = MulticoreParam(workers = 4)
my_bpparam <- MulticoreParam(workers = 4)
res2 <- SingleR(test = sce.mat,
ref = ref.mat,
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8 changes: 5 additions & 3 deletions episodes/eda_qc.Rmd
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Expand Up @@ -72,8 +72,8 @@ bcrank <- barcodeRanks(counts(sce))
# Only showing unique points for plotting speed.
uniq <- !duplicated(bcrank$rank)
line_df = data.frame(cutoff = names(metadata(bcrank)),
value = unlist(metadata(bcrank)))
line_df <- data.frame(cutoff = names(metadata(bcrank)),
value = unlist(metadata(bcrank)))
ggplot(bcrank[uniq,], aes(rank, total)) +
geom_point() +
Expand All @@ -92,6 +92,8 @@ A simple approach would be to apply a threshold on the total count to only retai

::: callout
Depending on your data source, identifying and discarding empty droplets may not be necessary. Some academic institutions have research cores dedicated to single cell work that perform the sample preparation and sequencing. Many of these cores will also perform empty droplet filtering and other initial QC steps. If the sequencing outputs were provided to you by someone else, make sure to communicate with them about what pre-processing steps have been performed, if any.

<!-- TODO: cite official 10x CellRanger docs -->
:::

:::: challenge
Expand Down Expand Up @@ -671,7 +673,7 @@ e.out <- emptyDrops(counts(sce))
sce <- sce[,which(e.out$FDR <= 0.001)]
# Thankfully the data come with gene symbols, which we can use to identify mitochondrial genes:
is.mito = grepl("^MT-", rowData(sce)$Symbol)
is.mito <- grepl("^MT-", rowData(sce)$Symbol)
# QC metrics ----
df <- perCellQCMetrics(sce, subsets = list(Mito = is.mito))
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2 changes: 1 addition & 1 deletion episodes/hca.Rmd
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Expand Up @@ -179,7 +179,7 @@ data.
For the sake of demonstration, we'll focus this small subset of samples:

```{r}
sample_subset = metadata |>
sample_subset <- metadata |>
filter(
ethnicity == "African" &
grepl("10x", assay) &
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6 changes: 3 additions & 3 deletions episodes/intro-sce.Rmd
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Expand Up @@ -234,9 +234,9 @@ The `SingleCellExperiment` constructor function can be used to create a new `Sin
::: solution

```{r}
mat = matrix(runif(30), ncol = 5)
mat <- matrix(runif(30), ncol = 5)
my_sce = SingleCellExperiment(assays = list(logcounts = mat))
my_sce <- SingleCellExperiment(assays = list(logcounts = mat))
my_sce$my_col_info = runif(5)
Expand All @@ -260,7 +260,7 @@ sce <- WTChimeraData(samples = 5)
sce6 <- WTChimeraData(samples = 6)
combined_sce = cbind(sce, sce6)
combined_sce <- cbind(sce, sce6)
combined_sce
```
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12 changes: 6 additions & 6 deletions episodes/large_data.Rmd
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Expand Up @@ -388,8 +388,8 @@ reducedDim(r.out, "PCA") = -1 * reducedDim(r.out, "PCA")
From there we can visualize the error with a histogram:

```{r}
error = reducedDim(r.out, "PCA")[,"PC1"] -
reducedDim(e.out, "PCA")[,"PC1"]
error <- reducedDim(r.out, "PCA")[,"PC1"] -
reducedDim(e.out, "PCA")[,"PC1"]
data.frame(approx_error = error) |>
ggplot(aes(approx_error)) +
Expand Down Expand Up @@ -569,15 +569,15 @@ function for writing to HDF5 from the `r Biocpkg("HDF5Array")` package.

```{r}
wt_out = tempfile(fileext = ".h5")
wt_out <- tempfile(fileext = ".h5")
wt_counts = counts(WTChimeraData())
wt_counts <- counts(WTChimeraData())
writeHDF5Array(wt_counts,
name = "wt_counts",
file = wt_out)
oom_wt = HDF5Array(wt_out, "wt_counts")
oom_wt <- HDF5Array(wt_out, "wt_counts")
object.size(wt_counts)
Expand Down Expand Up @@ -607,7 +607,7 @@ Use the function `system.time` to obtain the runtime of each job.

```{r eval=FALSE}
sce.brain = logNormCounts(sce.brain)
sce.brain <- logNormCounts(sce.brain)
system.time({i.out <- runPCA(sce.brain,
ncomponents = 20,
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