neoflow icon indicating copy to clipboard operation
neoflow copied to clipboard

Error with demo data when running STEP2

Open biosunsci opened this issue 3 years ago • 7 comments

I copied test data from example_data folder provided by the neoflow. the input_vcf_list.txt file looks like:

experiment      sample  file    file_type
crc_test        sample1 /neoantigen2/input/sample1.vcf   somatic

the copied sample1.vcf looks like:

##fileformat=VCFv4.2
##contig=<ID=chrM,length=16571,assembly=hg19>
##contig=<ID=chr1,length=249250621,assembly=hg19>
##contig=<ID=chr2,length=243199373,assembly=hg19>
##contig=<ID=chr3,length=198022430,assembly=hg19>
##contig=<ID=chr4,length=191154276,assembly=hg19>
##contig=<ID=chr5,length=180915260,assembly=hg19>
##contig=<ID=chr6,length=171115067,assembly=hg19>
##contig=<ID=chr7,length=159138663,assembly=hg19>
##contig=<ID=chr8,length=146364022,assembly=hg19>
##contig=<ID=chr9,length=141213431,assembly=hg19>
##contig=<ID=chr10,length=135534747,assembly=hg19>
##contig=<ID=chr11,length=135006516,assembly=hg19>
##contig=<ID=chr12,length=133851895,assembly=hg19>
##contig=<ID=chr13,length=115169878,assembly=hg19>
##contig=<ID=chr14,length=107349540,assembly=hg19>
##contig=<ID=chr15,length=102531392,assembly=hg19>
##contig=<ID=chr16,length=90354753,assembly=hg19>
##contig=<ID=chr17,length=81195210,assembly=hg19>
##contig=<ID=chr18,length=78077248,assembly=hg19>
##contig=<ID=chr19,length=59128983,assembly=hg19>
##contig=<ID=chr20,length=63025520,assembly=hg19>
##contig=<ID=chr21,length=48129895,assembly=hg19>
##contig=<ID=chr22,length=51304566,assembly=hg19>
##contig=<ID=chrX,length=155270560,assembly=hg19>
##contig=<ID=chrY,length=59373566,assembly=hg19>
##contig=<ID=chr1_gl000191_random,length=106433,assembly=hg19>
##contig=<ID=chr1_gl000192_random,length=547496,assembly=hg19>
##contig=<ID=chr4_gl000193_random,length=189789,assembly=hg19>
##contig=<ID=chr4_gl000194_random,length=191469,assembly=hg19>
##contig=<ID=chr6_cox_hap2,length=4795371,assembly=hg19>
##contig=<ID=chr7_gl000195_random,length=182896,assembly=hg19>
##contig=<ID=chr9_gl000198_random,length=90085,assembly=hg19>
##contig=<ID=chr9_gl000199_random,length=169874,assembly=hg19>
##contig=<ID=chr17_gl000205_random,length=174588,assembly=hg19>
##contig=<ID=chr19_gl000208_random,length=92689,assembly=hg19>
##contig=<ID=chr19_gl000209_random,length=159169,assembly=hg19>
##contig=<ID=chrUn_gl000211,length=166566,assembly=hg19>
##contig=<ID=chrUn_gl000212,length=186858,assembly=hg19>
##contig=<ID=chrUn_gl000213,length=164239,assembly=hg19>
##contig=<ID=chrUn_gl000214,length=137718,assembly=hg19>
##contig=<ID=chrUn_gl000215,length=172545,assembly=hg19>
##contig=<ID=chrUn_gl000216,length=172294,assembly=hg19>
##contig=<ID=chrUn_gl000217,length=172149,assembly=hg19>
##contig=<ID=chrUn_gl000218,length=161147,assembly=hg19>
##contig=<ID=chrUn_gl000219,length=179198,assembly=hg19>
##contig=<ID=chrUn_gl000220,length=161802,assembly=hg19>
##contig=<ID=chrUn_gl000221,length=155397,assembly=hg19>
##contig=<ID=chrUn_gl000222,length=186861,assembly=hg19>
##contig=<ID=chrUn_gl000224,length=179693,assembly=hg19>
##contig=<ID=chrUn_gl000225,length=211173,assembly=hg19>
##contig=<ID=chrUn_gl000226,length=15008,assembly=hg19>
##contig=<ID=chrUn_gl000228,length=129120,assembly=hg19>
##contig=<ID=chrUn_gl000229,length=19913,assembly=hg19>
##contig=<ID=chrUn_gl000230,length=43691,assembly=hg19>
##contig=<ID=chrUn_gl000231,length=27386,assembly=hg19>
##contig=<ID=chrUn_gl000232,length=40652,assembly=hg19>
##contig=<ID=chrUn_gl000234,length=40531,assembly=hg19>
##contig=<ID=chrUn_gl000236,length=41934,assembly=hg19>
##contig=<ID=chrUn_gl000237,length=45867,assembly=hg19>
##contig=<ID=chrUn_gl000238,length=39939,assembly=hg19>
##contig=<ID=chrUn_gl000239,length=33824,assembly=hg19>
##contig=<ID=chrUn_gl000240,length=41933,assembly=hg19>
##contig=<ID=chrUn_gl000241,length=42152,assembly=hg19>
##contig=<ID=chrUn_gl000243,length=43341,assembly=hg19>
##reference=file:///data/kail/tmp/nxf.ovMAqCSHCz/ucsc.hg19.fasta
#CHROM	POS	ID	REF	ALT	QUAL	FILTER	INFO	FORMAT	crc_cell_lines
chrM	73	.	G	A	207.78	PASS	.	GT:AD:DP:GQ:PL	1/1:0,9:9:27:236,27,0
chrM	150	.	T	C	85.28	PASS	.	GT:AD:DP:GQ:PL	1/1:0,3:3:9:113,9,0
chrM	410	.	A	T	169.84	PASS	.	GT:AD:DP:GQ:PL	1/1:0,6:6:18:198,18,0
chrM	515	.	GCA	G	83.25	PASS	.	GT:AD:DP:GQ:PL	1/1:0,3:3:9:120,9,0
chrM	2354	.	C	T	57.28	PASS	.	GT:AD:DP:GQ:PL	1/1:0,3:3:9:85,9,0
chrM	2708	.	G	A	126.9	PASS	.	GT:AD:DP:GQ:PL	1/1:0,5:5:15:155,15,0
chrM	16364	.	T	C	241.84	PASS	.	GT:AD:DP:GQ:PL	1/1:0,6:6:18:270,18,0
chr1	13868	.	A	G	72.77	PASS	.	GT:AD:DP:GQ:PL	0/1:16,6:22:99:101,0,367
chr1	13896	rs201696125	C	A	50.77	PASS	.	GT:AD:DP:GQ:PL	0/1:14,5:19:79:79,0,340
chr1	14907	rs79585140	A	G	919.77	PASS	.	GT:AD:DP:GQ:PL	0/1:20,38:58:99:948,0,429
chr1	14930	rs75454623	A	G	878.77	PASS	.	GT:AD:DP:GQ:PL	0/1:16,34:50:99:907,0,341
chr1	15274	rs201931625	A	T	62.28	PASS	.	GT:AD:DP:GQ:PL	1/1:0,3:3:9:90,9,0
chr1	16495	rs141130360	G	C	41.77	PASS	.	GT:AD:DP:GQ:PL	0/1:14,5:19:70:70,0,395
chr1	16534	rs201459529	C	T	298.77	PASS	.	GT:AD:DP:GQ:PL	0/1:5,15:20:67:327,0,67
chr1	16571	rs199676946	G	A	188.77	PASS	.	GT:AD:DP:GQ:PL	0/1:4,10:14:61:217,0,61
chr1	1453304	rs141065088	C	T	40.74	PASS	.	GT:AD:DP:GQ:PL	1/1:0,2:2:6:68,6,0
chr1	1454519	rs370448805	G	C	51.77	PASS	.	GT:AD:DP:GQ:PL	0/1:2,3:5:63:80,0,63
chr1	3427514	rs146093263	GCACACGCCCCCACC	G	98.25	PASS	.	GT:AD:DP:GQ:PL	1/1:0,3:3:9:135,9,0
chr1	3428160	rs2820999	T	G	215.8	PASS	.	GT:AD:DP:GQ:PL	1/1:0,7:7:21:244,21,0
chr1	3496479	rs2794340	T	C	33.77	PASS	.	GT:AD:DP:GQ:PL	0/1:2,2:4:62:62,0,64
chr1	3545175	rs147637374	GTTCTGGGAGCTCCTCCCCC	G	151.82	PASS	.	GT:AD:DP:GQ:PL	1/1:0,4:4:17:189,17,0
chr1	3546264	rs137965653	CCGGG	C	277.77	PASS	.	GT:AD:DP:GQ:PL	1/1:0,7:7:21:315,21,0
chr1	59147926	rs12139511	T	C	769.77	PASS	.	GT:AD:DP:GQ:PL	1/1:0,22:22:66:798,66,0
chr1	59148118	rs17118103	A	T	280.77	PASS	.	GT:AD:DP:GQ:PL	0/1:8,10:18:99:309,0,173
chr1	59150941	rs2206764	G	A	396.77	PASS	.	GT:AD:DP:GQ:PL	1/1:0,13:13:39:425,39,0
chr1	59150997	rs5774421	TA	T	88.87	PASS	.	GT:AD:DP:GQ:PL	1/1:0,5:5:15:126,15,0
chr1	59156155	rs12076049	C	T	200.84	PASS	.	GT:AD:DP:GQ:PL	1/1:0,6:6:18:229,18,0
chr1	59158632	rs12097333	T	C	584.77	PASS	.	GT:AD:DP:GQ:PL	1/1:0,18:18:54:613,54,0
chr1	59160724	rs66629118	TA	T	63.0	PASS	.	GT:AD:DP:GQ:PL	1/1:0,4:4:12:100,12,0
chr1	59248813	rs2760499	G	C	292.77	PASS	.	GT:AD:DP:GQ:PL	1/1:0,11:11:33:321,33,0
...(etc, 81914 lines in total)

I can run through STEP2 using these 2 files.

(base) [root@gpu01 neoantigen2]# sh test_cmd.sh 
N E X T F L O W  ~  version 21.04.3
Launching `./neoflow_db.nf` [compassionate_jennings] - revision: 421847b8f4
Format input mapping file: /neoantigen2/input/test_vcf_files.tsv!
New mapping file: output/test_vcf_files.tsv!
executor >  local (5)
[7c/318276] process > pre_processing (test_vcf_files.tsv)            [100%] 1 of 1 ✔
[87/d3b425] process > variant_annotation (crc_test-mapping_file.tsv) [100%] 1 of 1 ✔
[62/9f2138] process > database_construction (crc_test)               [100%] 1 of 1 ✔
[dd/0d806c] process > format_db (crc_test)                           [100%] 1 of 1 ✔
[05/178767] process > generate_decoy_db (crc_test)                   [100%] 1 of 1 ✔
Completed at: 19-Jan-2022 05:12:16
Duration    : 12m 59s
CPU hours   : 0.2
Succeeded   : 5


crc_test
N E X T F L O W  ~  version 21.04.3
Launching `./neoflow_msms.nf` [dreamy_borg] - revision: 5cf4143baf
Process multiple MS/MS files.
executor >  local (19)
[2b/f3de6e] process > msms_searching (spec-00037.mgf)                   [100%] 15 of 15 ✔
[d6/3db5c3] process > calculate_fdr (calculate_fdr)                     [100%] 1 of 1 ✔
[ab/67eada] process > prepare_pepquery_input (prepare_pepquery_input)   [100%] 1 of 1 ✔
[47/b207b0] process > run_pepquery (run_pepquery)                       [100%] 1 of 1 ✔
[3c/c1bbee] process > add_pepquery_validation (add_pepquery_validation) [100%] 1 of 1 ✔
Completed at: 19-Jan-2022 06:14:34
Duration    : 1h 2m 11s
CPU hours   : 11.0
Succeeded   : 19

but when I randomly sample 7191 vcf records from sample1.vcf before running the same pipeline, it is exactly the same as the last one except for the number of rows. this time the input vcf file looks like:

##fileformat=VCFv4.2
##contig=<ID=chrM,length=16571,assembly=hg19>
##contig=<ID=chr1,length=249250621,assembly=hg19>
##contig=<ID=chr2,length=243199373,assembly=hg19>
##contig=<ID=chr3,length=198022430,assembly=hg19>
##contig=<ID=chr4,length=191154276,assembly=hg19>
##contig=<ID=chr5,length=180915260,assembly=hg19>
##contig=<ID=chr6,length=171115067,assembly=hg19>
##contig=<ID=chr7,length=159138663,assembly=hg19>
##contig=<ID=chr8,length=146364022,assembly=hg19>
##contig=<ID=chr9,length=141213431,assembly=hg19>
##contig=<ID=chr10,length=135534747,assembly=hg19>
##contig=<ID=chr11,length=135006516,assembly=hg19>
##contig=<ID=chr12,length=133851895,assembly=hg19>
##contig=<ID=chr13,length=115169878,assembly=hg19>
##contig=<ID=chr14,length=107349540,assembly=hg19>
##contig=<ID=chr15,length=102531392,assembly=hg19>
##contig=<ID=chr16,length=90354753,assembly=hg19>
##contig=<ID=chr17,length=81195210,assembly=hg19>
##contig=<ID=chr18,length=78077248,assembly=hg19>
##contig=<ID=chr19,length=59128983,assembly=hg19>
##contig=<ID=chr20,length=63025520,assembly=hg19>
##contig=<ID=chr21,length=48129895,assembly=hg19>
##contig=<ID=chr22,length=51304566,assembly=hg19>
##contig=<ID=chrX,length=155270560,assembly=hg19>
##contig=<ID=chrY,length=59373566,assembly=hg19>
##contig=<ID=chr1_gl000191_random,length=106433,assembly=hg19>
##contig=<ID=chr1_gl000192_random,length=547496,assembly=hg19>
##contig=<ID=chr4_gl000193_random,length=189789,assembly=hg19>
##contig=<ID=chr4_gl000194_random,length=191469,assembly=hg19>
##contig=<ID=chr6_cox_hap2,length=4795371,assembly=hg19>
##contig=<ID=chr7_gl000195_random,length=182896,assembly=hg19>
##contig=<ID=chr9_gl000198_random,length=90085,assembly=hg19>
##contig=<ID=chr9_gl000199_random,length=169874,assembly=hg19>
##contig=<ID=chr17_gl000205_random,length=174588,assembly=hg19>
##contig=<ID=chr19_gl000208_random,length=92689,assembly=hg19>
##contig=<ID=chr19_gl000209_random,length=159169,assembly=hg19>
##contig=<ID=chrUn_gl000211,length=166566,assembly=hg19>
##contig=<ID=chrUn_gl000212,length=186858,assembly=hg19>
##contig=<ID=chrUn_gl000213,length=164239,assembly=hg19>
##contig=<ID=chrUn_gl000214,length=137718,assembly=hg19>
##contig=<ID=chrUn_gl000215,length=172545,assembly=hg19>
##contig=<ID=chrUn_gl000216,length=172294,assembly=hg19>
##contig=<ID=chrUn_gl000217,length=172149,assembly=hg19>
##contig=<ID=chrUn_gl000218,length=161147,assembly=hg19>
##contig=<ID=chrUn_gl000219,length=179198,assembly=hg19>
##contig=<ID=chrUn_gl000220,length=161802,assembly=hg19>
##contig=<ID=chrUn_gl000221,length=155397,assembly=hg19>
##contig=<ID=chrUn_gl000222,length=186861,assembly=hg19>
##contig=<ID=chrUn_gl000224,length=179693,assembly=hg19>
##contig=<ID=chrUn_gl000225,length=211173,assembly=hg19>
##contig=<ID=chrUn_gl000226,length=15008,assembly=hg19>
##contig=<ID=chrUn_gl000228,length=129120,assembly=hg19>
##contig=<ID=chrUn_gl000229,length=19913,assembly=hg19>
##contig=<ID=chrUn_gl000230,length=43691,assembly=hg19>
##contig=<ID=chrUn_gl000231,length=27386,assembly=hg19>
##contig=<ID=chrUn_gl000232,length=40652,assembly=hg19>
##contig=<ID=chrUn_gl000234,length=40531,assembly=hg19>
##contig=<ID=chrUn_gl000236,length=41934,assembly=hg19>
##contig=<ID=chrUn_gl000237,length=45867,assembly=hg19>
##contig=<ID=chrUn_gl000238,length=39939,assembly=hg19>
##contig=<ID=chrUn_gl000239,length=33824,assembly=hg19>
##contig=<ID=chrUn_gl000240,length=41933,assembly=hg19>
##contig=<ID=chrUn_gl000241,length=42152,assembly=hg19>
##contig=<ID=chrUn_gl000243,length=43341,assembly=hg19>
##reference=file:///data/kail/tmp/nxf.ovMAqCSHCz/ucsc.hg19.fasta
#CHROM	POS	ID	REF	ALT	QUAL	FILTER	INFO	FORMAT	crc_cell_lines
chrM	73	.	G	A	207.78	PASS	.	GT:AD:DP:GQ:PL	1/1:0,9:9:27:236,27,0
chrM	150	.	T	C	85.28	PASS	.	GT:AD:DP:GQ:PL	1/1:0,3:3:9:113,9,0
chrM	410	.	A	T	169.84	PASS	.	GT:AD:DP:GQ:PL	1/1:0,6:6:18:198,18,0
chrM	515	.	GCA	G	83.25	PASS	.	GT:AD:DP:GQ:PL	1/1:0,3:3:9:120,9,0
chrM	2354	.	C	T	57.28	PASS	.	GT:AD:DP:GQ:PL	1/1:0,3:3:9:85,9,0
chrM	2708	.	G	A	126.9	PASS	.	GT:AD:DP:GQ:PL	1/1:0,5:5:15:155,15,0
chrM	16364	.	T	C	241.84	PASS	.	GT:AD:DP:GQ:PL	1/1:0,6:6:18:270,18,0
chr1	3546264	rs137965653	CCGGG	C	277.77	PASS	.	GT:AD:DP:GQ:PL	1/1:0,7:7:21:315,21,0
chr1	59465926	rs2764901	A	C	337.77	PASS	.	GT:AD:DP:GQ:PL	0/1:22,11:33:99:366,0,1040
chr1	60280749	rs113730234	GT	G	360.74	PASS	.	GT:AD:DP:GQ:PL	1/1:0,10:10:30:398,30,0
chr1	62231924	rs141761262	T	TTGA	53.7	PASS	.	GT:AD:DP:GQ:PL	1/1:0,2:2:6:90,6,0
chr1	62627675	rs2481676	G	A	191.77	PASS	.	GT:AD:DP:GQ:PL	0/1:6,7:13:99:220,0,170
chr1	63113511	rs746735	C	A	202.77	PASS	.	GT:AD:DP:GQ:PL	0/1:4,7:11:99:231,0,133
chr1	64097432	rs1126728	C	T	273.77	PASS	.	GT:AD:DP:GQ:PL	0/1:13,10:23:99:302,0,360
chr1	64698411	rs2029868	T	C	45.77	PASS	.	GT:AD:DP:GQ:PL	0/1:1,2:3:29:74,0,29
chr1	67288045	rs482082	C	T	140.77	PASS	.	GT:AD:DP:GQ:PL	0/1:8,6:14:99:169,0,245
chr1	67562106	rs6670381	G	T	46.74	PASS	.	GT:AD:DP:GQ:PL	1/1:0,2:2:6:74,6,0
chr1	70397087	rs7530457	T	C	54.77	PASS	.	GT:AD:DP:GQ:PL	0/1:7,3:10:83:83,0,233
...(etc, 7191 lines in total)

I got an fatal Error when running the last procedure of STEP2 whose Error message is :

executor >  local (19)                                                                                              [60/1956]
[a3/308b14] process > msms_searching (spec-00037.mgf)                   [100%] 15 of 15 ✔
[ae/c73260] process > calculate_fdr (calculate_fdr)                     [100%] 1 of 1 ✔
[c4/516371] process > prepare_pepquery_input (prepare_pepquery_input)   [100%] 1 of 1 ✔
[68/fd6ea1] process > run_pepquery (run_pepquery)                       [100%] 1 of 1 ✔
[67/fd2463] process > add_pepquery_validation (add_pepquery_validation) [100%] 1 of 1, failed: 1 ✘
Error executing process > 'add_pepquery_validation (add_pepquery_validation)'

Caused by:
  Process `add_pepquery_validation (add_pepquery_validation)` terminated with an error exit status (1)

Command executed:

  #!/usr/bin/env /usr/local/bin/Rscript
     library(dplyr)
     library(readr)
     library(stringr)
  
     psms <- read_tsv("novel_peptides_psm.tsv")
  
     psm_rank_file = "pepquery/psm_rank.txt"
     if(file.exists(psm_rank_file)){
         psm_rank <- read_tsv(psm_rank_file)
  if("n_ptm" %in% names(psm_rank)){
             psm_rank <- psm_rank %>% filter(pvalue<=0.01,n_ptm==0,rank==1)
             psms$pepquery <- ifelse(psms$peptide %in% psm_rank$peptide,1,0)
  }else{
      psms$pepquery <- 0ter(),
  }
     }else{
         psms$pepquery <- 0
     }
     psms %>% write_tsv("novel_peptides_psm_pepquery.tsv")

Command exit status:
  1

Command output:
  (empty)

Command error:
  Bioconductor version 3.10 (BiocManager 1.30.10), ?BiocManager::install for help
  Bioconductor version '3.10' is out-of-date; the current release version '3.14'
    is available with R version '4.1'; see https://bioconductor.org/install
  
  Attaching package: ‘dplyr’
  
  The following objects are masked from ‘package:stats’:
 
      filter, lag
  
  The following objects are masked from ‘package:base’:
  
      intersect, setdiff, setequal, union
  
  Parsed with column specification:
  cols(
    index = col_character(),
    evalue = col_character(),
    ion_score = col_character(),
    charge = col_character(),
    mass = col_character(),
    mz = col_character(),
    delta_da = col_character(),
    delta_ppm = col_character(),
    peptide = col_character(),
    isdecoy = col_character(),
    miss = col_character(),
    protein = col_character(),
    position = col_character(),
    rt = col_character(),
    isSAP = col_character(),
    mods = col_character(),
    FDR = col_character(),
    Qvalue = col_character(),
    is_novel = col_character()
  )
  Error in `$<-.data.frame`(`*tmp*`, pepquery, value = 0) : 
    replacement has 1 row, data has 0
  Calls: $<- -> $<-.data.frame
  Execution halted

Work dir:
 /neoantigen2/work/67/fd2463881a197b5022a22c8c2c78a2

Tip: when you have fixed the problem you can continue the execution adding the option `-resume` to the run command line

I think it is caused by the empty of the intermediate file: /neoantigen2/work/7d/3269b5449820da00611a534f20e697/novel_peptides_psm.tsv and the novel_peptides_psm.tsv looks like:

index   evalue  ion_score       charge  mass    mz      delta_da        delta_ppm       peptide isdecoy miss    protein posit

I don't understand why the number of rows affect the pipeline's execution.

Could you help me out ?

biosunsci avatar Jan 19 '22 06:01 biosunsci

@bingzhang16 Could you please help me?

biosunsci avatar Jan 19 '22 07:01 biosunsci

Could you send me the second vcf file (7191 records)?

wenbostar avatar Jan 19 '22 15:01 wenbostar

Could you send me the second vcf file (7191 records)?

sample7796.vcf.zip please unzip it then you'll get the vcf file I listed. I'm sorry about the wrong number of lines I counted, but it still can trigger the error I mentioned.

I've tried to sample the data to different number of rows from 500 ~ 10000, all the same error. @wenbostar

biosunsci avatar Jan 20 '22 03:01 biosunsci

Could you share with me the command line you used for both step 1 and step 2?

wenbostar avatar Jan 25 '22 18:01 wenbostar

Could you share with me the command line you used for both step 1 and step 2?

I'm using the sh test_cmd.sh to test step 1 and step2 and the content of the test_cmd.sh is

test_cmd.sh

docker run --rm -it  -w neoantigen2 --user root -v /seq:/seq -v /var/run/docker.sock:/var/run/docker.sock bzhanglab/neoflow:1.0   ./nextflow run ./neoflow_db.nf --ref_dir ./annovar/humandb_hg19/ --vcf_file ./input/test_vcf_files.tsv --annovar_dir ./annovar/ --ref_ver hg19 --out_dir output --cpu 30 &&
 echo crc_test && docker run --rm -it  -w neoantigen2 --user root -v /seq:/seq -v /var/run/docker.sock:/var/run/docker.sock bzhanglab/neoflow:1.0   ./nextflow run ./neoflow_msms.nf     --ms refs/mgf/     --msms_para_file refs/comet_parameter.txt     --search_engine comet     --db output/customized_database/crc_test_target_decoy.fasta     --out_dir output_crc_test/     --pv_refdb output/customized_database/ref.fasta     --pv_tol 20   --pv_itol 0.05  --prefix crc_test  &&  echo done!

The other dir structures or file contents are listed below in case to help.

ls -la /seq/neoantigen2

drwxr-xr-x 1 1024 983 33 Jan 12 11:35 ./ drwxrwxr-x 1 1024 1026 17 Jan 5 05:20 ../ drwxr-xr-x 1 1024 983 7 Jan 4 01:27 annovar/ drwxr-xr-x 1 1024 983 8 Jan 4 01:27 bin/ drwxr-xr-x 1 1024 983 1 Jan 4 01:27 database/ drwxr-xr-x 1 1024 983 11 Jan 12 09:16 input/ -rwxr-xr-x 1 1024 983 6636 Jan 4 01:27 neoflow_db.nf* -rwxr-xr-x 1 1024 983 3193 Jan 4 01:27 neoflow_hlatyping.nf* -rwxr-xr-x 1 1024 983 10976 Jan 4 01:27 neoflow_msms.nf* -rwxr-x--- 1 1024 983 8774 Jan 4 01:27 neoflow_neoantigen.nf* drwxr-xr-x 1 1024 983 6 Jan 4 01:27 netMHCpan-4.0/ drwxr-xr-x 1 root root 3 Jan 12 12:12 .nextflow/ -rwxr-xr-x 1 1024 983 56633890 Jan 4 01:27 nextflow* -rwxr-xr-x 1 1024 983 142 Jan 4 01:27 nextflow.config* drwxr-xr-x 1 root root 3 Jan 12 10:25 output/ drwxr-xr-x 1 root root 3 Jan 12 11:03 output_crc_test/ **drwxr-xr-x 1 1024 983 2 Jan 4 01:27 refs/ -rw-r--r-- 1 root root 911 Jan 13 01:55 test_cmd.sh drwxr-xr-x 1 root root 190 Jan 12 12:12 work/

ls -la ./input/

drwxr-xr-x 1 1024 983 11 Jan 12 09:16 ./ drwxr-xr-x 1 1024 983 33 Jan 12 11:35 ../ -rw-r--r-- 1 root root 55571 Jan 12 08:04 500.txt -rw-r--r-- 1 root root 46592 Jan 12 08:04 noGQ-PL-500.txt -rw-r--r-- 1 root root 5115845 Jan 12 07:55 noGQ-PL.txt -rw-r--r-- 1 root root 6068515 Jan 11 10:03 noGQ.txt -rw-r--r-- 1 root root 5573011 Jan 11 10:04 noPL.txt -rw-r--r-- 1 root root 6557121 Jan 13 01:57 sample1.txt -rw-r--r-- 1 1024 983 6557121 Jan 13 01:58 sample1.vcf -rw-r--r-- 1 1024 983 127 Jan 13 03:01 test_vcf_files.tsv

 less ./input/test_vcf_files.tsv
experiment      sample  file    file_type
crc_test        sample1 /seq/neoantigen2/input/500.txt    somatic
 tree ./refs
./refs
├── ./comet_parameter.txt
└── ./mgf/
    ├── spec-00030.mgf
    ├── spec-00139.mgf
    ├── spec-00279.mgf
    ├── spec-00364.mgf
    ├── spec-00560.mgf
    ├── spec-00037.mgf
    ├── spec-00208.mgf
    ├── spec-00285.mgf
    ├── spec-00373.mgf
    ├── spec-00565.mgf
    ├── spec-00129.mgf
    ├── spec-00219.mgf
    ├── spec-00351.mgf
    ├── spec-00465.mgf
    └── spec-00622.mgf

biosunsci avatar Jan 30 '22 04:01 biosunsci

@wenbostar I've pasted the relative information, Could you find any clues about the error I met?

biosunsci avatar Feb 07 '22 04:02 biosunsci

Hello, @biosunsci Unlike the command you executed, I am not using docker to run the script, just locally, and all the required software and docker in NeoFlow have been successfully installed locally. But I never managed to run step 2, and I got the error #9. So I was very excited when I saw that you were able to run step 2. According to your above description in #3 , I guess maybe I should use docker to run example. However, I am not clear whether I need to create a docker and install software such as an novar and nextflow in it? I have never used docker before😶‍🌫️, and I really hope you can give me some advice, please~

Sawakotx avatar May 16 '23 12:05 Sawakotx