Comments (9)
Hi Biao, I agree that the current log files does not suggest that the issue is caused by different strands, nevertheless, given that different strands is the only known source for causing this error, could you please use fixchr and check whether that solves the issue. Also, please share the dotplots generated by fixchr.
from syri.
Thanks for replying.
Instead of using fixchr , I used chroder to tune the chr name and chain direction of the original sequence. The command line is as follows
nucmer -c 1000 -l 40 -t 16 -p TGY ref.fa TGY.fa
delta-filter -1 -i 90 -l 100 TGY.delta > TGY.1delta
show-coords -THrd TGY.1delta > TGY.1coords
chroder -o TGY TGY.1coords ref.fa TGY.fa
Then, I ran the above codes.
Actually, I analyzed a total of six sequence files, four of which succeeded and two of which failed. I double-checked the format of these files but did not find the problem. After reading the issues in your forum, I guess it might be a problem that chromosomes are from different strands.
I will run it again with fixchr.
from syri.
I ran
fixchr --prefix TGY -c TGY.1coords -r ref.fa -q TGY.qry.fasta
(2023-06-01 02:35:01,134 - fixchr - WARNING - Inverting query chromosomes: ['Chr07'] (fixchr.py:77))
output files:
TGYinput_alignments.txt
TGYinput.pdf
TGYhomologous_alignments.txt
TGYhomologous.pdf
TGYhomologous_strand_corrected_alignments.txt
TGYhomologous_strand_corrected.pdf
ref.filtered.fa
TGY.qry.filtered.fa
TGYinput.pdf
TGYhomologous_strand_corrected.pdf
Should I use result file TGY.qry.filtered.fa for subsequent analysis?
from syri.
Yes, try using that for alignment and then running syri.
from syri.
Hi, Manish
I performed the two files (TGY and DASZ) that did not fail to analyze before in the above way. TGY one ran successfully, but strangely the DASZ one still reported the same error log.
output files:
DASZinput.pdf
DASZhomologous_alignments.txt
DASZhomologous.pdf
DASZhomologous_strand_corrected_alignments.txt
DASZhomologous_strand_corrected.pdf
And the log file:
DASZ.syri.log
DASZinput.pdf
DASZhomologous_strand_corrected.pdf
DASZhomologous.pdf
Could please help me
DASZ.syri.log
analyze the reason again?
Many thanks
from syri.
It seems that fixchr
incorrectly reverse complemented chromosome 5 (compare DASZinput.pdf vs DASZhomologous_strand_corrected.pdf) . Using the original strand should solve the issue. You can use the dotplot
tool (installed with fixchr) to visualise and ensure the chromosomes are collinear.
from syri.
I used the original strand of Chr05, but the whole run analysis still didn't work. The generated intermediate files are as follows:
DASZ.syri.log
So, I tried to invert all the chromosomes without output content (Chr05, 06, 13,14,15) again and run it again, but it still failed.
I used the dotplot tool to visualize the linearity of these chromosomes, but the run failed with the following feedback:
"ValueError: Length mismatch: Expected axis has 11 elements, new values have 12 elements"
Then, I started the processing and analysis of the data of DASZ sample all over again. However, it still failed (even though I reversed Chr05 again).
input.pdf
homologous_strand_corrected.pdf
I don't have a clue anymore. Could please help me analyze the reason again?
from syri.
The log files show that some chromosomes have high fraction of inverted alignments, so yes there still seems to be issues with the strands. Also, I noticed that the Chromosome Ids are not consistent (ex: Chr11 in the two genomes are not homologous, and Chr4 and Chr3 are named interchangeably). This again can result in crashes as syri cannot find syntenic regions between homologous chromosomes (chromosomes with same ID). Could you please rename the chromosomes so that homologous chromosomes have same ID? After that try to ensure that you do not get warnings like this: 2023-06-09 11:34:25,376 - Reading Coords - WARNING - syri:135 - Reference chromosome Chr11 has high fraction of inverted alignments with its homologous chromosome in the query genome (Chr11). Ensure that same chromosome-strands are being compared in the two genomes, as different strand can result in unexpected errors.
If it still crashes then you can try to compare homologous chromosomes individually with syri (Chr1vsChr1, Chr2vsChr2 etc). That might help in better pinpointing the cause of the issue.
from syri.
Thanks Manish.
I successfully ran the analysis of this file after splitting this genome into separate runs for each stain file to check for problems!
All tasks have been successfully completed.
Many thanks~
from syri.
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