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explain scan/diff params.
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LKremer authored Oct 25, 2024
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Expand Up @@ -168,7 +168,7 @@ Now that `filtered_data` is smoothed, we can proceed with the VMR detection:
methscan scan --threads 4 filtered_data VMRs.bed
```
We use the option `--threads 4` in order to run the program on 4 CPU threads in parallel. If you want to use all available threads, simply omit the `--threads` option altogether.
The result is a [BED-file](https://en.wikipedia.org/wiki/BED_(file_format)) that lists the genomic coordinates (chromosome, start, end) of regions where methylation is variable between cells, as well as the methylation variance of the region:
The result is a [BED](https://en.wikipedia.org/wiki/BED_(file_format))-like file that lists the genomic coordinates (chromosome, start, end) of regions where methylation is variable between cells, as well as the methylation variance of the region:
```
2 3194798 3197978 0.07718534715010719
2 3379038 3381638 0.08048814475349723
Expand All @@ -178,7 +178,6 @@ The result is a [BED-file](https://en.wikipedia.org/wiki/BED_(file_format)) that
...
```


### 4. Obtaining a methylation matrix

Finally, you can quantify the mean methylation of the VMRs that we just discovered using `methscan matrix`:
Expand Down Expand Up @@ -414,6 +413,9 @@ methscan diff --threads 4 filtered_data cell_groups.csv DMRs.bed
The output file `DMRs.bed` contains a list of DMRs, their genome coordinates, the methylation difference measured by the t-statistic, and an adjusted p-value for each DMR.
One way to explore potential functions of these DMRs is to use tools such as [GREAT](http://great.stanford.edu).

Of course you can tweak the parameters of both `scan` and `diff` to your needs. If you are primarily interested in large stretches of differentially methylated DNA, for instance, you can increase the bandwidth of the sliding window and/or use `--bridge-gaps` to merge small VMRs/DMRs that are very close.
<img src="tutorial_scan_params.png" height="300">



### Considerations for real data sets
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