diff --git a/docs/content/counts-nano.md b/docs/content/counts-nano.md new file mode 100644 index 00000000..b071ba77 --- /dev/null +++ b/docs/content/counts-nano.md @@ -0,0 +1,104 @@ +# counts-nano - compute single-site methylation from nanopore data + +## Synopsis +```console +$ dnmtools counts-nano [OPTIONS] -c +``` + +## Description + +The `counts-nano` command introduced in v1.5.0 is designed specifically to +generate DNMTools [counts](../counts) format files from nanopore data called +for the `5mCG_5hmCG` modification. Currently this is only supported for +methylation and hydroxymethylation called at CpG sites. + +More documentation will come as this tool evolves, but for now: + +- Most behavior is very similar to what you will find from [counts](../counts). +- Mutation information is not estimated by `nano-counts`. +- Currently this only works for CpG sites and when the only modified sites are + marked as `C+m?` or `C+h?` in the `MM` field of each BAM/SAM read record. +- The first 6 columns of the output are the same as explained in the + [counts](../counts) format, except the fraction for the 5th column is both + 5mC and 5hmC. The 7th column is for 5hmC alone and the 8th is for 5mC alone. +- The methylation levels will not result in integer values when multiplied by + the number of reads because probabilities on modifications are used, so + methylation levels for each site are expected values (the best estimates we + can make), and do not use arbitrary cutoffs. +- Other commands in DNMTools have been modified to use this form of expected + methylation level, and behave as previously for bisulfite sequencing data, + but have updated behavior when the data is from nanopore. The user does not + need to specify the technology used. +- Some commands need to use a `-relaxed` flag to work with the additional + columns in the output from `counts-nano` compared with `counts`. For + commands without this option, simply do `cut -f1-6` on the output of + `counts-nano` to remove those. + +## Options + +```txt +-o, -output +``` +Output file name. The default is to write output to the terminal, +which is not useful unless commands are piped. + +```txt +-c, -chrom +``` +Reference genome file, which must be in FASTA format. This is +required. + +```txt +-t, -threads +``` + +The number of threads to use. This is only really helpful if the input is BAM +(not helpful for SAM), and the output is to be zipped (see `-z` below). These +threads are used to decompress BAM input and compress gzip output. If only one +of these conditions holds, using more threads can still help. Because most +computation in `counts-nano` is processing reads sequentially, using too many +threads will have decreasing returns. + +```txt +-z, -zip +``` + +The output should be zipped (in gzip format). This is not deduced by the +filename, but specifying this argument should be accompanied by using a `.gz` +filename suffix for the output. + +```txt +-n, -cpg-only +``` + +Print only CpG context cytosines. This significantly reduces the output size +in most genomes. Note that using this option does not merge data as symmetric +CpGs. + +```txt +-sym +``` + +This will turn on `-n, -cpg-only` automatically and will output symmetric CpG +sites, with each level including all counts and methylation levels as a +(weighted) average of both strands. + +```txt +-H, -header +``` + +Add a header to the output file to identify the reference genome. This will be +in the form of "comment" lines beginning with `#`. This is not required for most +downstream processing, but is used by commands that check for consistency with +a reference genome. + +```txt +-v, -verbose +``` + +Report more information while the program is running. + +```txt +-progress +``` +Show progress while the program is running.