Skip to content

Commit

Permalink
Version 1.10.0
Browse files Browse the repository at this point in the history
   * Output N barcodes per subdirectory with `--files-per-directory N` and output splitting
   * BioSample awareness for XML input and split output and allow ignoring them with `--ignore-biosamples`
   * Increase `--window-size-mult` to `3` to allow longer spacers
   * Do not report no adapter hits as too short inserts
   * Increase `--guess` barcode score to `75` if `--peek-guess --ccs` are combined
   * Enable double demux of CCS data
   * Print run time, CPU time, and peak memory consumption with `--log-level INFO`
   * New CLI UX
  • Loading branch information
armintoepfer committed Sep 9, 2019
1 parent c02cc7e commit 082883e
Showing 1 changed file with 37 additions and 3 deletions.
40 changes: 37 additions & 3 deletions README.md
Original file line number Diff line number Diff line change
Expand Up @@ -115,7 +115,7 @@ to use `--no-pbi`, omit the pbi index file, to minimize time to result.

## Input data
Input data is either raw unaligned subreads, straight from a Sequel, or
unaligned CCS reads, generated by [CCS 2](https://github.com/PacificBiosciences/unanimity);
unaligned CCS reads, generated by [CCS](https://github.com/PacificBiosciences/ccs);
both in the PacBio enhanced BAM format. If you want to demux RSII data, first
use SMRT Link or bax2bam to convert h5 to BAM. In addition, a `datastore.json`
with one file entry, either a SubreadSet or ConsensusReadSet, is also allowed.
Expand Down Expand Up @@ -462,7 +462,7 @@ there is no way to distinguish between pairs `bc1001--bc1002` and `bc1002--bc100
Score and tag per subread, instead per ZMW.

### `--window-size-mult`
The candidate region size multiplier: `barcode_length * multiplier`, default `1.5`.
The candidate region size multiplier: `barcode_length * multiplier`, default `3`.
Optionally, you can specify the region size in base pairs with `--window-size-bp`.

### Alignment options
Expand Down Expand Up @@ -986,9 +986,43 @@ false positives.
* Calls barcodes per barcode region and does not enforce adapter coupling
* Nice reports for QC

### Can I remove PCR primers after demultiplexing?
Yes! After demultiplexing, just lima on the output again with your PCR primer(s).

### Can I limit the output files per directory?
If you use output BAM splitting, it can happen that you get a lot of output files.
Using `--files-per-directory N` creates subdirectories and outputs at most `N`
barcodes per directory.

### `--peek-guess` does not work with XML input!
If your input XML file contains `<BioSamples>`, lima will deactivate barcode
inference via `--peek-guess` and only output barcodes specified in this section.
The assumption is that you know exactly which barcodes have been used and need no
inference. If this assumption is wrong, like the barcodes in the XML are wrong,
you can either just use BAM as input or use `--ignore-biosamples`.

### Help, I get `ERROR: Could not find matching barcodes!`
If you happen to get following error message

ERROR: Could not find matching barcodes! Check that the set of barcodes contains the used sequences and the correct mode has been selected: same or different.

then your XML input contains BioSamples with different barcode names than the
provided `barcode.fasta` file. Please check that you've used the correct
barcodes. You can ignore barcodes specified in the XML with `--ignore-biosamples`.


## Full Changelog

* **1.9.0**:
* **1.10.0**:
* Output N barcodes per subdirectory with `--files-per-directory N` and output splitting
* BioSample awareness for XML input and split output and allow ignoring them with `--ignore-biosamples`
* Increase `--window-size-mult` to `3` to allow longer spacers
* Do not report no adapter hits as too short inserts
* Increase `--guess` barcode score to `75` if `--peek-guess --ccs` are combined
* Enable double demux of CCS data
* Print run time, CPU time, and peak memory consumption with `--log-level INFO`
* New CLI UX
* 1.9.0:
* Add `--bad-adapter-ratio` to remove ZMWs with molecularly missing adapters
* Fix rare case, where a read only matches one barcode and not a single alternative
* Fix `--no-bam` to automatically omit pbi
Expand Down

0 comments on commit 082883e

Please sign in to comment.