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chen42 avatar chen42 commented on August 19, 2024 1

@lcoombe I tried several parameters. For my dataset, increasing e to 9000 has the largest effect on N50, when a is between 0.7 and 0.9. Thank you very much for your help in getting my error sorted out! I am closing this ticket.

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lcoombe avatar lcoombe commented on August 19, 2024

Hi @chen42,
Have you been able to successfully run the test example (Examples/arcs_test-demo)?
Could you provide all of the commands that were run when you used the arcs-make Makefile?
Does your tigpair_checkpoint.tsv file have pairs of contigs listed? (Feel free to include a few lines of the file here if you aren't sure)

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chen42 avatar chen42 commented on August 19, 2024

Hi, @lcoombe,
Thank you very much for looking into my problem!

  1. I installed arc in my homedir on a cluster. There is not enough diskspace in my home to run the test. So I copied the Example scripts to a project directory and ran the runARCSdemo.sh command and it did finish and produced the scaffold.fa file. I just checked the error log and found the following message:
Traceback (most recent call last):
  File "./makeTSVfile.py", line 96, in <module>
    readGraphFile(infile)
  File "./makeTSVfile.py", line 15, in readGraphFile
    with open(infile, 'r') as f:
FileNotFoundError: [Errno 2] No such file or directory: 'hsapiens-8reformat.fa.scaff_s98_c5_l0_d0_e30000_r0.05_original.gv'

The tigpair_checkpoint.tsv file is empty.

  1. For my own data, my arcs-make script is very simple:
cd /lustre/haven/proj/UTHSC0013/tigmint_data/arcs/
./arcs-make arcs draft=rn6_after_tigment reads=bnmale_barcode t=80
  1. here are several lines of my tigpair_checkpoint.tsv:
10      r1021   r1661   1       100
10      f1661   f1021   1       100
10      r1021   r3226   1       100
10      f3226   f1021   1       100
10      f1025   f1037   1       100
10      r1037   r1025   1       100
10      f103    r1823   1       100
10      f1823   r103    1       100
10      f1057   f959    1       100
10      r959    r1057   1       100
....
....
10      r931    f935    1       100
10      r935    f931    1       100
10      r9495   r9536   1       100
10      f9536   f9495   1       100
10      f953    r955    1       100
10      f955    r953    1       100
10      f981    f989    1       100
10      r989    r981    1       100

Thanks!

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lcoombe avatar lcoombe commented on August 19, 2024

Thanks @chen42,
For your own run, the fact that the tigpair checkpoint file isn't empty is a good sign - that means that some connections are being made, although the ones that you are showing aren't very supported - only 1 barcode supports each of those links. Are all the values in column 4 of your tigpair_checkpoint tsv 1?

Can you also post a few lines of your reads file, to just be doubly sure that the read headers are formatted correctly?

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chen42 avatar chen42 commented on August 19, 2024

@lcoombe: I found the problem. My reads.fq.gz file was corrupted. So when I checked the file with zcat, the first few lines were OK but there were not many reads in the file. The result is much better after fixing that issue, although most values in col 4 of the tigpair_checkpoint.tsv is still 1:

$ cut -f 4 rn6_after_tigment_c5_m50-10000_s98_r0.05_e30000_z500.tigpair_checkpoint.tsv |sort |uniq -c |sort -rn |head
  54662 1
    398 2
    208 3
    174 4
    168 6
    168 5
    154 7
    150 8
    140 10
    134 9

Trying with a more accurate range for m:

$ cut -f 4 rn6_after_tigment_c5_m50-3000_s98_r0.05_e30000_z500.tigpair_checkpoint.tsv |sort |uniq -c |sort -rn |head
  54344 1
    398 2
    208 3
    178 4
    168 6
    164 5
    154 7
    150 8
    146 10
    132 9

I assume the value of a does not affect this. Any suggestion on which parameter I should try next? Thank you!

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chen42 avatar chen42 commented on August 19, 2024

I forgot to provide the first few lines of my reads file:

@E00247:397:HJ7YLALXX:8:1101:12195:19768_AAACACCAGCGATATA
TATACCACACACACACACCCCACACCATACCACACATATACCGCACAAACACACACCCCCCCCCCCATCCCGCACCGCTATCCCATATGCCCACCCCTGCCCGCCACTCATCCCCCGACCACACTTCA
+
FJJJJFAFFJJFJJ<J<JJJJJJJJJ<JJFFFFJJJFJJFAJ--A7---<FJ-<-7----7---7--77-7-A7------7A-A-77--------)7)))))))-)))7----7))))-))7------
@E00247:397:HJ7YLALXX:8:1101:12195:19768_AAACACCAGCGATATA
ATGTGTGGTATGGTGTGGGGTGTGTGTGTGTGGGATACTCGCAGTATATCGCTGGTGTTGAGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTAGATCTCGGTGGTCGCCGTATCATTAAAAAGTAAGTGAGAGGTGGTGGGTGGTTGCGG
+
AAFAFJJJJJJJJJJJJJFJJJJJJJJJJJJJA--<A-<-----7<7-AAFA-7F7A---7777-<A-7F<F-7--AAAFFFJF-A7F-F<AA-A-<<JAFJ<AJ<)<-)A))----7--7-7----------77-)77)7-)))7--))7
@E00247:397:HJ7YLALXX:8:1101:2798:21456_AAACACCAGCGATATA
CCCACACACACACACAACACACAACACACACACACACAGGCACATACACACACACAACACACAGACACAACACACACAACACACACATACACCCACCACACACACACACAGCACACACAACACCCCCC
+
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJFJJFJJJJJ<JJJJJJFJJJFJJFJJJ-77FAJ<FJAJ-A7A7AJAAAF-F<-<F<AJFFFJ)FAF<FFJAJ<F--7<J7<-A--A-A-7--)

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lcoombe avatar lcoombe commented on August 19, 2024

@chen42 - Glad to hear you worked out the issues!

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