Closed IsmailM closed 5 years ago
Changing the below to check against None
rather than an empty string (""
) fixes the issue for me:
Does this repo still maintained ? I have the same issue, and I am wondering if this has been addressed.
Yes, it is maintained. I will check this problem
Simon
On Tue, 8 Jan 2019, 09:11 Anand Mayakonda <notifications@github.com wrote:
Does this repo still maintained ? I have the same issue, and I am wondering if this has been addressed.
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Hi Simon, Thanks for the reply. Here is the error log I have ..
:
: Command map started at 2019-01-08 10:18:43.945975
:
: ------------ Mapping Parameters ------------
: Sample barcode : AS-277115-LR-38819
: Data set : AS-277115-LR-38819
: No. threads : 2
: Index : /bigdisk/ref_db/mm10/gemBS/mm10.BS.gem
: Paired : True
: Read non stranded: True
: Type : PAIRED
: Input Files : AS-277115-LR-38819_R1.fastq.gz,AS-277115-LR-38819_R2.fastq.gz
: Output dir : /mapping/AS-277115-LR-38819
:
: Bisulfite Mapping...
Traceback (most recent call last):
File "/usr/local/bin/gemBS", line 11, in <module>
load_entry_point('gemBS==3.2.2', 'console_scripts', 'gemBS')()
File "/usr/local/lib/python3.5/dist-packages/gemBS/commands.py", line 157, in gemBS_main
instances[args.command].run(args)
File "/usr/local/lib/python3.5/dist-packages/gemBS/production.py", line 367, in run
self.do_mapping(fl)
File "/usr/local/lib/python3.5/dist-packages/gemBS/production.py", line 559, in do_mapping
under_conversion=self.underconversion_sequence,over_conversion=self.overconversion_sequence)
File "/usr/local/lib/python3.5/dist-packages/gemBS/__init__.py", line 737, in mapping
process = run_tools(tools, name="bisulfite-mapping", logfile=logfile)
File "/usr/local/lib/python3.5/dist-packages/gemBS/utils.py", line 330, in run_tools
p.start()
File "/usr/local/lib/python3.5/dist-packages/gemBS/utils.py", line 237, in start
logging.info("Starting:\n\t%s" % (self.to_bash_pipe()))
File "/usr/local/lib/python3.5/dist-packages/gemBS/utils.py", line 276, in to_bash_pipe
return " | ".join([p.to_bash() for p in self.processes])
File "/usr/local/lib/python3.5/dist-packages/gemBS/utils.py", line 276, in <listcomp>
return " | ".join([p.to_bash() for p in self.processes])
File "/usr/local/lib/python3.5/dist-packages/gemBS/utils.py", line 165, in to_bash
return " ".join(self.commands)
TypeError: sequence item 17: expected str instance, NoneType found
I pushed a bug fix to github - can you check to see if this fixes your problem?
Thanks, Simon
On 8 Jan 2019, at 09:21, Anand Mayakonda notifications@github.com wrote:
Hi Simon, Thanks for the reply. Here is the error log I have ..
: : Command map started at 2019-01-08 10:18:43.945975 : : ------------ Mapping Parameters ------------ : Sample barcode : AS-277115-LR-38819 : Data set : AS-277115-LR-38819 : No. threads : 2 : Index : /bigdisk/ref_db/mm10/gemBS/mm10.BS.gem : Paired : True : Read non stranded: True : Type : PAIRED : Input Files : AS-277115-LR-38819_R1.fastq.gz,AS-277115-LR-38819_R2.fastq.gz : Output dir : /mapping/AS-277115-LR-38819 : : Bisulfite Mapping... Traceback (most recent call last): File "/usr/local/bin/gemBS", line 11, in
load_entry_point('gemBS==3.2.2', 'console_scripts', 'gemBS')() File "/usr/local/lib/python3.5/dist-packages/gemBS/commands.py", line 157, in gemBS_main instances[args.command].run(args) File "/usr/local/lib/python3.5/dist-packages/gemBS/production.py", line 367, in run self.do_mapping(fl) File "/usr/local/lib/python3.5/dist-packages/gemBS/production.py", line 559, in do_mapping under_conversion=self.underconversion_sequence,over_conversion=self.overconversion_sequence) File "/usr/local/lib/python3.5/dist-packages/gemBS/init.py", line 737, in mapping process = run_tools(tools, name="bisulfite-mapping", logfile=logfile) File "/usr/local/lib/python3.5/dist-packages/gemBS/utils.py", line 330, in run_tools p.start() File "/usr/local/lib/python3.5/dist-packages/gemBS/utils.py", line 237, in start logging.info("Starting:\n\t%s" % (self.to_bash_pipe())) File "/usr/local/lib/python3.5/dist-packages/gemBS/utils.py", line 276, in to_bash_pipe return " | ".join([p.to_bash() for p in self.processes]) File "/usr/local/lib/python3.5/dist-packages/gemBS/utils.py", line 276, in return " | ".join([p.to_bash() for p in self.processes]) File "/usr/local/lib/python3.5/dist-packages/gemBS/utils.py", line 165, in to_bash return " ".join(self.commands) TypeError: sequence item 17: expected str instance, NoneType found — You are receiving this because you commented. Reply to this email directly, view it on GitHub https://github.com/heathsc/gemBS/issues/37#issuecomment-452212300, or mute the thread https://github.com/notifications/unsubscribe-auth/ADHPdx09zroRqqNLXElkIW50uUuoPtDRks5vBFT9gaJpZM4XAo1t.
Hi Simon, Thanks for the quick fix. Alignment went smoothly, however I have new error in BScall.. Below is the log..
$ gemBS call
:
: Command call started at 2019-01-08 12:24:02.381098
:
: ----------- Methylation Calling --------
: Reference : /bigdisk/scJens/gem_bs/genome/mm10.fa
: Species : mm10
: Right Trim : 10
: Left Trim : 10
: Chromosomes : ['chr1', 'chr2', 'chrX', 'chr3', 'chr4', 'chr5', 'chr6', 'chr7', 'chr10', 'chr8', 'chr14', 'chr9', 'chr11', 'chr13', 'chr12', 'chr15', 'chr16', 'chr17', 'chrY', 'chr18', 'chr19', '@pool_1']
: Threads : 6
: Sample: AS-277115-LR-38819 Bam: /bigdisk/scJens/gem_bs//mapping/AS-277115-LR-38819/AS-277115-LR-38819.bam
:
: Methylation Calling...
2019-01-08 12:25:06,267 ERROR: Process '/home/epicwl/miniconda3/lib/python3.6/site-packages/gemBS/gemBSbinaries/bs_call' finished with 1
2019-01-08 12:25:06,267 ERROR: Loading reference sequences
2019-01-08 12:25:06,267 ERROR: Completed loading reference sequences
2019-01-08 12:25:06,267 ERROR: Processing chromosome chr2 (OK)
2019-01-08 12:25:06,267 ERROR: > System.Error::Signal raised (no=11)
Exception in thread Thread-2:
Traceback (most recent call last):
ValueError: Error while executing the bscall process.
And the above error just hangs, I had to terminate the process.
OK, this is not good! Anyway I can get access to the files so that I can try and reproduce the error on my system?
Simon
On 8 Jan 2019, at 11:30, Anand Mayakonda notifications@github.com wrote:
Hi Simon, Thanks for the quick fix. Alignment went smoothly, however I have new error in BScall.. Below is the log..
: : Command call started at 2019-01-08 12:24:02.381098 : : ----------- Methylation Calling -------- : Reference : /bigdisk/scJens/gem_bs/genome/mm10.fa : Species : mm10 : Right Trim : 10 : Left Trim : 10 : Chromosomes : ['chr1', 'chr2', 'chrX', 'chr3', 'chr4', 'chr5', 'chr6', 'chr7', 'chr10', 'chr8', 'chr14', 'chr9', 'chr11', 'chr13', 'chr12', 'chr15', 'chr16', 'chr17', 'chrY', 'chr18', 'chr19', '@pool_1'] : Threads : 6 : Sample: AS-277115-LR-38819 Bam: /bigdisk/scJens/gem_bs//mapping/AS-277115-LR-38819/AS-277115-LR-38819.bam : : Methylation Calling... 2019-01-08 12:25:06,267 ERROR: Process '/home/epicwl/miniconda3/lib/python3.6/site-packages/gemBS/gemBSbinaries/bs_call' finished with 1 2019-01-08 12:25:06,267 ERROR: Loading reference sequences 2019-01-08 12:25:06,267 ERROR: Completed loading reference sequences 2019-01-08 12:25:06,267 ERROR: Processing chromosome chr2 (OK) 2019-01-08 12:25:06,267 ERROR: > System.Error::Signal raised (no=11) Exception in thread Thread-2: Traceback (most recent call last): ValueError: Error while executing the bscall process. And the above error just hangs, I had to terminate the process.
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Do you need fastq files or the bam file ?
Also this is a single cell library with non-directional protocol. I quickly did map-report and it says 33% overall alignment rate before deduplication (attached). For same sample when I used Bismark it reported ~18% alignment.
While you take a look at the error, could you suggest me if I can use any downstream tools deduplication (is picard fine?) and for methylation extraction ?
Just the BAM should be fine. Which reference are you using? Normally deduplication is not required (it is performed by the caller). If you could send a copy of the parameters files you are using for gemBS that would be helpful.
Thanks,
Simon
On 8 Jan 2019, at 11:45, Anand Mayakonda notifications@github.com wrote:
Do you need fastq files or the bam file ?
Also this is a single cell library with non-directional protocol. I quickly did map-report and it says 33% overall alignment rate before deduplication (attached). For same sample when I used Bismark it reported ~18% alignment.
AS-277115-LR-38819.html.zip https://github.com/heathsc/gemBS/files/2736357/AS-277115-LR-38819.html.zip While you take a look at the error, could you suggest me if I can use any downstream tools deduplication (is picard fine?) and for methylation extraction ?
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Could you please let me know once you download the file so that I can remove it from the Dropbox. Thank you and hope this helps. Also is it ok to use MethylDackel for extraction ?
OK, I have the files. I’ll let you know how I get on with testing.
I have no experience with MethylDackel. As far as I can see it does not take account of any possible sequence variation so the output should be treated with caution. I’m not sure how well it will work with BAMs generated from gemBS - it depends whether it recognizes the tags in the BAM file that describe the bisulfite strand. All bisulfite mappers use different tags for this (bs_call understands the BAMs from about 5 different mappers).
Simon
On 8 Jan 2019, at 12:23, Anand Mayakonda notifications@github.com wrote:
Here https://www.dropbox.com/s/047fjs49teb1zmc/AS-277115-LR-38819.bam?dl=0 scbs_samples.conf.zip https://github.com/heathsc/gemBS/files/2736506/scbs_samples.conf.zip Could you please let me know once you download the file so that I can remove it from the Dropbox. Thank you and hope this helps. Also is it ok to use MethylDackel for extraction ?
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Ah! Got it. Thank you.
The calling for your files runs without problems on my system. How did you install gemBS? Would it be possible to try the Docker or Singularity containers to see if these work for you?
Simon
On 8 Jan 2019, at 13:48, Anand Mayakonda notifications@github.com wrote:
Ah! Got it. Thank you.
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I followed the instructions as in the docs for installation.
git clone --recursive https://github.com/heathsc/gemBS.git
python3 setup.py install
I do not have experience with docker/singularity but I see there are instructions in the docs. I will try it and let you know.
Thank you for the help.
Should be OK then - what type of computer are you running this on (CPU, memory, OS etc.) ?
Simon
On 8 Jan 2019, at 16:15, Anand Mayakonda notifications@github.com wrote:
I followed the instructions as in the docs for installation.
git clone --recursive https://github.com/heathsc/gemBS.git python3 setup.py install I do not have experience with docker/singularity but I see there are instructions in the docs. I will try it and let you know.
Thank you for the help.
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I am running it on a local system with 128G RAM and 20 threads running on Ubuntu 16.04 LTS. I just installed it on another system, will let you know if I face the same issuse..
That should be more than enough and Ubuntu 16.04 is a tested system. If you re-run the calling with less threads does it make a difference (it shouldn’t do, but I’m trying to pinpoint what could be triggering the bug).
Simon
On 8 Jan 2019, at 16:37, Anand Mayakonda notifications@github.com wrote:
I am running it on a local system with 128G RAM and 20 threads running on Ubuntu 16.04 LTS. I just installed it on another system, will let you know if I face the same issuse..
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Okay, I will check. Should I start with the alignment ? Or is there a way to use just 1 thread for gemBS call
, since the config file has been set to 6 threads.
Just the calling. Maybe set it to 1 thread to see if it runs through.
Simon
On 8 Jan 2019, at 16:44, Anand Mayakonda notifications@github.com wrote:
Okay, I will check. Should I start with the alignment ? Or is there a way to use just 1 thread for gemBS call, since the config file has been set to 6 threads.
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Nope, still the same issue. I changed threads valueto 1 in .gemBS/gemBS.json
and ran the command.
gemBS call
:
: Command call started at 2019-01-08 18:00:30.938804
:
: ----------- Methylation Calling --------
: Reference : /bigdisk/scJens/gem_bs/genome/mm10.fa
: Species : mm10
: Right Trim : 10
: Left Trim : 10
: Chromosomes : ['chr1', 'chr2', 'chrX', 'chr3', 'chr4', 'chr5', 'chr6', 'chr7', 'chr10', 'chr8', 'chr14', 'chr9', 'chr11', 'chr13', 'chr12', 'chr15', 'chr16', 'chr17', 'chrY', 'chr18', 'chr19', '@pool_1']
: Threads : 1
: Sample: AS-277115-LR-38819 Bam: /bigdisk/scJens/gem_bs//mapping/AS-277115-LR-38819/AS-277115-LR-38819.bam
:
: Methylation Calling...
2019-01-08 18:01:08,477 ERROR: Process '/home/epicwl/miniconda3/lib/python3.6/site-packages/gemBS/gemBSbinaries/bs_call' finished with 1
2019-01-08 18:01:08,478 ERROR: Loading reference sequences
2019-01-08 18:01:08,478 ERROR: Completed loading reference sequences
2019-01-08 18:01:08,478 ERROR: Processing chromosome chr7 (OK)
2019-01-08 18:01:08,478 ERROR: > System.Error::Signal raised (no=11)
Exception in thread Thread-2:
Traceback (most recent call last):
ValueError: Error while executing the bscall process.
And this happens straightaway after the line 'Processing chromosome chr7 (OK)’ is printed?
Simon
On 8 Jan 2019, at 17:04, Anand Mayakonda notifications@github.com wrote:
Nope, still the same issue. I changed threads valueto 1 in .gemBS/gemBS.json and ran the command.
gemBS call : : Command call started at 2019-01-08 18:00:30.938804 : : ----------- Methylation Calling -------- : Reference : /bigdisk/scJens/gem_bs/genome/mm10.fa : Species : mm10 : Right Trim : 10 : Left Trim : 10 : Chromosomes : ['chr1', 'chr2', 'chrX', 'chr3', 'chr4', 'chr5', 'chr6', 'chr7', 'chr10', 'chr8', 'chr14', 'chr9', 'chr11', 'chr13', 'chr12', 'chr15', 'chr16', 'chr17', 'chrY', 'chr18', 'chr19', '@pool_1'] : Threads : 1 : Sample: AS-277115-LR-38819 Bam: /bigdisk/scJens/gem_bs//mapping/AS-277115-LR-38819/AS-277115-LR-38819.bam : : Methylation Calling... 2019-01-08 18:01:08,477 ERROR: Process '/home/epicwl/miniconda3/lib/python3.6/site-packages/gemBS/gemBSbinaries/bs_call' finished with 1 2019-01-08 18:01:08,478 ERROR: Loading reference sequences 2019-01-08 18:01:08,478 ERROR: Completed loading reference sequences 2019-01-08 18:01:08,478 ERROR: Processing chromosome chr7 (OK) 2019-01-08 18:01:08,478 ERROR: > System.Error::Signal raised (no=11) Exception in thread Thread-2: Traceback (most recent call last): ValueError: Error while executing the bscall process. — You are receiving this because you commented. Reply to this email directly, view it on GitHub https://github.com/heathsc/gemBS/issues/37#issuecomment-452353095, or mute the thread https://github.com/notifications/unsubscribe-auth/ADHPd0g6KAS6hUVgndwv3CUUJycB8w_Mks5vBMGbgaJpZM4XAo1t.
Its weird. Every time I run gemBS call
it stops at the next chromosome.
: Methylation Calling...
2019-01-08 18:10:16,045 ERROR: Process '/home/epicwl/miniconda3/lib/python3.6/site-packages/gemBS/gemBSbinaries/bs_call' finished with 1
2019-01-08 18:10:16,045 ERROR: Loading reference sequences
2019-01-08 18:10:16,045 ERROR: Completed loading reference sequences
2019-01-08 18:10:16,045 ERROR: Processing chromosome chr10 (OK)
2019-01-08 18:10:16,046 ERROR: > System.Error::Signal raised (no=11)
Exception in thread Thread-1:
Traceback (most recent call last):
ValueError: Error while executing the bscall process.
2019-01-08 18:10:16,083 ERROR: Process '/home/epicwl/miniconda3/lib/python3.6/site-packages/gemBS/gemBSbinaries/bs_call' finished with 1
2019-01-08 18:10:16,083 ERROR: Loading reference sequences
2019-01-08 18:10:16,083 ERROR: Completed loading reference sequences
2019-01-08 18:10:16,083 ERROR: Processing chromosome chr8 (OK)
2019-01-08 18:10:16,083 ERROR: > System.Error::Signal raised (no=11)
Exception in thread Thread-2:
Traceback (most recent call last):
ValueError: Error while executing the bscall process.
: Methylation call done, samples performed: AS-277115-LR-38819
I also think its still using more than one thread. How do I force it to use single thread ?
Is it completing the chromosomes each time? If you look in the calling directory, do you see a bcf file and a json file for the chromosome (particularly the json file, as this is only generated at the end)?
On 8 Jan 2019, at 17:12, Anand Mayakonda notifications@github.com wrote:
Its weird. Every time I run gemBS call it stops at the next chromosome.
: Methylation Calling... 2019-01-08 18:10:16,045 ERROR: Process '/home/epicwl/miniconda3/lib/python3.6/site-packages/gemBS/gemBSbinaries/bs_call' finished with 1 2019-01-08 18:10:16,045 ERROR: Loading reference sequences 2019-01-08 18:10:16,045 ERROR: Completed loading reference sequences 2019-01-08 18:10:16,045 ERROR: Processing chromosome chr10 (OK) 2019-01-08 18:10:16,046 ERROR: > System.Error::Signal raised (no=11) Exception in thread Thread-1: Traceback (most recent call last): ValueError: Error while executing the bscall process.
2019-01-08 18:10:16,083 ERROR: Process '/home/epicwl/miniconda3/lib/python3.6/site-packages/gemBS/gemBSbinaries/bs_call' finished with 1 2019-01-08 18:10:16,083 ERROR: Loading reference sequences 2019-01-08 18:10:16,083 ERROR: Completed loading reference sequences 2019-01-08 18:10:16,083 ERROR: Processing chromosome chr8 (OK) 2019-01-08 18:10:16,083 ERROR: > System.Error::Signal raised (no=11) Exception in thread Thread-2: Traceback (most recent call last): ValueError: Error while executing the bscall process.
: Methylation call done, samples performed: AS-277115-LR-38819
I also think its still using more than one thread. How do I force it to use single thread ?
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Yes, there are json and bcf files for the ones that are completed. But all the json files are empty.
Okay, on a different system it works fine and calling completed without any error. I guess it was an installation problem on the main system. Thank you for your patient replies and apologies for the mess :| Should have checked it properly.
Thanks for the feedback, but a segfault is never good and I would be happier knowing what caused it! If you get around to re-installing gemBS on the original system, please let me know if it works (and definitely if it still does not work!).
Simon
On 8 Jan 2019, at 17:46, Anand Mayakonda notifications@github.com wrote:
Okay, on a different system it works fine and calling completed without any error. I guess it was an installation problem on the main system. Thank you for your patient replies and apologies for the mess :| Should have checked it properly.
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Sure. Tomorrow I will check again and update you. Thanks again.
Hi Simon, I reinstalled everything on the new system and it worked fine. Still dont know how it messed up the first installation.
I have another question to ask. How do I run gemBS map with a custom input json file ?
$gemBS prepare -c ${conf} -t ${csv} --output ${sample_name}.json --no-db
$gemBS map
gemBS.utils.CommandException: gemBS JSON file not found.
How do I pass output json from prepare
command to map command ?
Thanks again.
To pass a custom json file, use the -j command line option before the subcommand so:
gemBS -j myfile.json map
Simon
On 9 Jan 2019, at 15:35, Anand Mayakonda notifications@github.com wrote:
Hi, Simon, I reinstalled everything on the new system and it worked fine. Still dont know how it messed up the first installation.
I have another question to ask. How do I run gemBS map with a custom input json file ?
$gemBS prepare -c ${conf} -t ${csv} --output ${sample_name}.json --no-db $gemBS map gemBS.utils.CommandException: gemBS JSON file not found. How do I pass output json from prepare command to map command ?
Thanks again.
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Ah! Nice. It works. I was using the wrong way then gemBS map -j myfile.json
Thanks again.
Thanks for fixing this @heathsc 👍
I see this error when trying to run
gemBS map
:Looking at this at more details - this issue is raised at the following location (specifically at L165):
https://github.com/heathsc/gemBS/blob/87a6657ac1055311d9f429c8237329912cbca723/gemBS/utils.py#L161-L166
In my case,
self.command
is equal to the following:As such it seems the issue because the values of
--underconversion_sequence
and--overconversion_sequence
is set toNone
.Thus it seems that there should be a default value (of an empty string) set for both of these parameters (or rather they shouldn't be added to this list if their value is
None
)