Skip to content

Commit

Permalink
Try to create space on GHA to avoid running out of space like at http…
Browse files Browse the repository at this point in the history
  • Loading branch information
lcolladotor committed May 21, 2024
1 parent 7c3f1f9 commit bf959c2
Showing 1 changed file with 15 additions and 1 deletion.
16 changes: 15 additions & 1 deletion vignettes/recount-workflow.Rmd
Original file line number Diff line number Diff line change
Expand Up @@ -729,7 +729,7 @@ recount2 provides bigWig coverage files (unscaled) for all samples, as well as a

For illustrative purposes, we will use the data from chromosome 21 for the SRP045638 project. First, we obtain the expressed regions using a relatively high mean cutoff of 5. We then filter the regions to keep only the ones longer than 100 base-pairs to shorten the time needed for running `coverage_matrix()`.

```{r 'download_bigwigs', eval = .Platform$OS.type != 'windows'}
```{r 'download_mean_bigwig', eval = .Platform$OS.type != 'windows'}
## Normally, one can use rtracklayer::import() to access remote parts of BigWig
## files without having to download the complete files. However, as of
## 2024-05-20 this doesn't seem to be working well. So this is a workaround to
Expand All @@ -754,10 +754,21 @@ table(width(regions) >= 100)
## Keep only the ones that are at least 100 bp long
regions <- regions[width(regions) >= 100]
length(regions)
## Remove file we no longer need
unlink("SRP045638/bw", recursive = TRUE)
```

Now that we have a set of regions to work with, we proceed to build a _RangedSummarizedExperiment_ object with the coverage counts, add the expanded metadata we built for the gene-level, and scale the counts. Note that `coverage_matrix()` scales the base-pair coverage counts by default, which we turn off in order to use use `scale_counts()`.

```{r 'download_sample_bigwigs', eval = .Platform$OS.type != 'windows'}
## Normally, one can use rtracklayer::import() to access remote parts of BigWig
## files without having to download the complete files. However, as of
## 2024-05-20 this doesn't seem to be working well. So this is a workaround to
## issue https://github.com/lawremi/rtracklayer/issues/83
download_study("SRP045638", type = "samples")
```

```{r "build_rse_ER", eval = .Platform$OS.type != "windows"}
## Compute coverage matrix for study SRP045638, only for chromosome 21
## Takes about 4 minutes
Expand All @@ -774,6 +785,9 @@ rse_er_scaled <- scale_counts(rse_er)
## To highlight that we scaled the counts
rm(rse_er)
## Remove files we no longer need
unlink("SRP045638/bw", recursive = TRUE)
```

Now that we have a scaled count matrix for the expressed regions, we can proceed with the DE analysis just like we did at the gene and exon feature levels (Figures \@ref(fig:erdeanalysis1), \@ref(fig:erdeanalysis2), \@ref(fig:erdeanalysis3), and \@ref(fig:erdeanalysis4)).
Expand Down

0 comments on commit bf959c2

Please sign in to comment.