nadegeguiglielmoni / GraphUnzip

Unzip assembly graphs with Hi-C data and/or long reads.
GNU General Public License v3.0
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GraphUnzip crashed #12

Closed jflot closed 2 years ago

jflot commented 2 years ago

I tried running GraphUnzip on a SPAdes Illumina assembly of a ca. 250 Mb genome, after aligning 156 000 Nanopore reads > 10 kb in length on the GFA using SPAligner (command: spaligner spaligner_config.yaml -d nanopore -s ONT_10kbmin.fastq -g SPADES_nanopore_assembly_graph_with_scaffolds.gfa -k 77 -t 14 -o spaligner. Alignment went well, with > 99.9% of reads mapping on the graph. However, GraphUnzip crashed afterward (command: graphunzip.py -g SPADES_assembly_graph_with_scaffolds.gfa -l spaligner/alignment.tsv -o SPADES_assembly_graph_with_scaffolds_GraphUnzip.gfa -f SPADES_assembly_graph_with_scaffolds_GraphUnzip.fasta unzip). Here is the log:

Loading the GFA file Loading contigs Loading links Everything loaded, moving on to untangling the graph Reading the gpa file... Finished going through the gpa file. Inventoried 1000 long reads over 73428 Inventoried 2000 long reads over 73428 Inventoried 3000 long reads over 73428 Inventoried 4000 long reads over 73428 Inventoried 5000 long reads over 73428 Inventoried 6000 long reads over 73428 Inventoried 7000 long reads over 73428 Inventoried 8000 long reads over 73428 Inventoried 9000 long reads over 73428 Inventoried 10000 long reads over 73428 Inventoried 11000 long reads over 73428 Inventoried 12000 long reads over 73428 Inventoried 13000 long reads over 73428 Inventoried 14000 long reads over 73428 Inventoried 15000 long reads over 73428 Inventoried 16000 long reads over 73428 Inventoried 17000 long reads over 73428 Inventoried 18000 long reads over 73428 Inventoried 19000 long reads over 73428 Inventoried 20000 long reads over 73428 Inventoried 21000 long reads over 73428 Inventoried 22000 long reads over 73428 Inventoried 23000 long reads over 73428 Inventoried 24000 long reads over 73428 Inventoried 25000 long reads over 73428 Inventoried 26000 long reads over 73428 Inventoried 27000 long reads over 73428 Inventoried 28000 long reads over 73428 Inventoried 29000 long reads over 73428 Inventoried 30000 long reads over 73428 Inventoried 31000 long reads over 73428 Inventoried 32000 long reads over 73428 Inventoried 33000 long reads over 73428 Inventoried 34000 long reads over 73428 Inventoried 35000 long reads over 73428 Inventoried 36000 long reads over 73428 Inventoried 37000 long reads over 73428 Inventoried 38000 long reads over 73428 Inventoried 39000 long reads over 73428 Inventoried 40000 long reads over 73428 Inventoried 41000 long reads over 73428 Inventoried 42000 long reads over 73428 Inventoried 43000 long reads over 73428 Inventoried 44000 long reads over 73428 Inventoried 45000 long reads over 73428 Inventoried 46000 long reads over 73428 Inventoried 47000 long reads over 73428 Inventoried 48000 long reads over 73428 Inventoried 49000 long reads over 73428 Inventoried 50000 long reads over 73428 Inventoried 51000 long reads over 73428 Inventoried 52000 long reads over 73428 Inventoried 53000 long reads over 73428 Inventoried 54000 long reads over 73428 Inventoried 55000 long reads over 73428 Inventoried 56000 long reads over 73428 Inventoried 57000 long reads over 73428 Inventoried 58000 long reads over 73428 Inventoried 59000 long reads over 73428 Inventoried 60000 long reads over 73428 Inventoried 61000 long reads over 73428 Inventoried 62000 long reads over 73428 Inventoried 63000 long reads over 73428 Inventoried 64000 long reads over 73428 Inventoried 65000 long reads over 73428 Inventoried 66000 long reads over 73428 Inventoried 67000 long reads over 73428 Inventoried 68000 long reads over 73428 Inventoried 69000 long reads over 73428 Inventoried 70000 long reads over 73428 Inventoried 71000 long reads over 73428 Inventoried 72000 long reads over 73428 Inventoried 73000 long reads over 73428 Building consensus bridges from all the long reads consensused 0 bridges out of 51462 consensused 100 bridges out of 51462 consensused 200 bridges out of 51462 consensused 300 bridges out of 51462 consensused 400 bridges out of 51462 consensused 500 bridges out of 51462 consensused 600 bridges out of 51462 consensused 700 bridges out of 51462 consensused 800 bridges out of 51462 consensused 900 bridges out of 51462 consensused 1000 bridges out of 51462 consensused 1100 bridges out of 51462 consensused 1200 bridges out of 51462 consensused 1300 bridges out of 51462 consensused 1400 bridges out of 51462 consensused 1500 bridges out of 51462 consensused 1600 bridges out of 51462 consensused 1700 bridges out of 51462 consensused 1800 bridges out of 51462 consensused 1900 bridges out of 51462 consensused 2000 bridges out of 51462 consensused 2100 bridges out of 51462 consensused 2200 bridges out of 51462 consensused 2300 bridges out of 51462 consensused 2400 bridges out of 51462 consensused 2500 bridges out of 51462 consensused 2600 bridges out of 51462 consensused 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out of 51462 consensused 5400 bridges out of 51462 consensused 5500 bridges out of 51462 consensused 5600 bridges out of 51462 consensused 5700 bridges out of 51462 consensused 5800 bridges out of 51462 consensused 5900 bridges out of 51462 consensused 6000 bridges out of 51462 consensused 6100 bridges out of 51462 consensused 6200 bridges out of 51462 consensused 6300 bridges out of 51462 consensused 6400 bridges out of 51462 consensused 6500 bridges out of 51462 consensused 6600 bridges out of 51462 consensused 6700 bridges out of 51462 consensused 6800 bridges out of 51462 consensused 6900 bridges out of 51462 consensused 7000 bridges out of 51462 consensused 7100 bridges out of 51462 consensused 7200 bridges out of 51462 consensused 7300 bridges out of 51462 consensused 7400 bridges out of 51462 consensused 7500 bridges out of 51462 consensused 7600 bridges out of 51462 consensused 7700 bridges out of 51462 consensused 7800 bridges out of 51462 consensused 7900 bridges out of 51462 consensused 8000 bridges out of 51462 consensused 8100 bridges out of 51462 consensused 8200 bridges out of 51462 consensused 8300 bridges out of 51462 consensused 8400 bridges out of 51462 consensused 8500 bridges out of 51462 consensused 8600 bridges out of 51462 consensused 8700 bridges out of 51462 consensused 8800 bridges out of 51462 consensused 8900 bridges out of 51462 consensused 9000 bridges out of 51462 consensused 9100 bridges out of 51462 consensused 9200 bridges out of 51462 consensused 9300 bridges out of 51462 consensused 9400 bridges out of 51462 consensused 9500 bridges out of 51462 consensused 9600 bridges out of 51462 consensused 9700 bridges out of 51462 consensused 9800 bridges out of 51462 consensused 9900 bridges out of 51462 consensused 10000 bridges out of 51462 consensused 10100 bridges out of 51462 consensused 10200 bridges out of 51462 consensused 10300 bridges out of 51462 consensused 10400 bridges out of 51462 consensused 10500 bridges out of 51462 consensused 10600 bridges out of 51462 consensused 10700 bridges out of 51462 consensused 10800 bridges out of 51462 consensused 10900 bridges out of 51462 consensused 11000 bridges out of 51462 consensused 11100 bridges out of 51462 consensused 11200 bridges out of 51462 consensused 11300 bridges out of 51462 consensused 11400 bridges out of 51462 consensused 11500 bridges out of 51462 consensused 11600 bridges out of 51462 consensused 11700 bridges out of 51462 consensused 11800 bridges out of 51462 consensused 11900 bridges out of 51462 consensused 12000 bridges out of 51462 consensused 12100 bridges out of 51462 consensused 12200 bridges out of 51462 consensused 12300 bridges out of 51462 consensused 12400 bridges out of 51462 consensused 12500 bridges out of 51462 consensused 12600 bridges out of 51462 consensused 12700 bridges out of 51462 consensused 12800 bridges out of 51462 consensused 12900 bridges out of 51462 consensused 13000 bridges out of 51462 consensused 13100 bridges 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out of 51462 consensused 26000 bridges out of 51462 consensused 26100 bridges out of 51462 consensused 26200 bridges out of 51462 consensused 26300 bridges out of 51462 consensused 26400 bridges out of 51462 consensused 26500 bridges out of 51462 consensused 26600 bridges out of 51462 consensused 26700 bridges out of 51462 consensused 26800 bridges out of 51462 consensused 26900 bridges out of 51462 consensused 27000 bridges out of 51462 consensused 27100 bridges out of 51462 consensused 27200 bridges out of 51462 consensused 27300 bridges out of 51462 consensused 27400 bridges out of 51462 consensused 27500 bridges out of 51462 consensused 27600 bridges out of 51462 consensused 27700 bridges out of 51462 consensused 27800 bridges out of 51462 consensused 27900 bridges out of 51462 consensused 28000 bridges out of 51462 consensused 28100 bridges out of 51462 consensused 28200 bridges out of 51462 consensused 28300 bridges out of 51462 consensused 28400 bridges out of 51462 consensused 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out of 51462 consensused 38800 bridges out of 51462 consensused 38900 bridges out of 51462 consensused 39000 bridges out of 51462 consensused 39100 bridges out of 51462 consensused 39200 bridges out of 51462 consensused 39300 bridges out of 51462 consensused 39400 bridges out of 51462 consensused 39500 bridges out of 51462 consensused 39600 bridges out of 51462 consensused 39700 bridges out of 51462 consensused 39800 bridges out of 51462 consensused 39900 bridges out of 51462 consensused 40000 bridges out of 51462 consensused 40100 bridges out of 51462 consensused 40200 bridges out of 51462 consensused 40300 bridges out of 51462 consensused 40400 bridges out of 51462 consensused 40500 bridges out of 51462 consensused 40600 bridges out of 51462 consensused 40700 bridges out of 51462 consensused 40800 bridges out of 51462 consensused 40900 bridges out of 51462 consensused 41000 bridges out of 51462 consensused 41100 bridges out of 51462 consensused 41200 bridges out of 51462 consensused 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bridges out of 51462 consensused 46500 bridges out of 51462 consensused 46600 bridges out of 51462 consensused 46700 bridges out of 51462 consensused 46800 bridges out of 51462 consensused 46900 bridges out of 51462 consensused 47000 bridges out of 51462 consensused 47100 bridges out of 51462 consensused 47200 bridges out of 51462 consensused 47300 bridges out of 51462 consensused 47400 bridges out of 51462 consensused 47500 bridges out of 51462 consensused 47600 bridges out of 51462 consensused 47700 bridges out of 51462 consensused 47800 bridges out of 51462 consensused 47900 bridges out of 51462 consensused 48000 bridges out of 51462 consensused 48100 bridges out of 51462 consensused 48200 bridges out of 51462 consensused 48300 bridges out of 51462 consensused 48400 bridges out of 51462 consensused 48500 bridges out of 51462 consensused 48600 bridges out of 51462 consensused 48700 bridges out of 51462 consensused 48800 bridges out of 51462 consensused 48900 bridges out of 51462 consensused 49000 bridges out of 51462 consensused 49100 bridges out of 51462 consensused 49200 bridges out of 51462 consensused 49300 bridges out of 51462 consensused 49400 bridges out of 51462 consensused 49500 bridges out of 51462 consensused 49600 bridges out of 51462 consensused 49700 bridges out of 51462 consensused 49800 bridges out of 51462 consensused 49900 bridges out of 51462 consensused 50000 bridges out of 51462 consensused 50100 bridges out of 51462 consensused 50200 bridges out of 51462 consensused 50300 bridges out of 51462 consensused 50400 bridges out of 51462 consensused 50500 bridges out of 51462 consensused 50600 bridges out of 51462 consensused 50700 bridges out of 51462 consensused 50800 bridges out of 51462 consensused 50900 bridges out of 51462 consensused 51000 bridges out of 51462 consensused 51100 bridges out of 51462 consensused 51200 bridges out of 51462 consensused 51300 bridges out of 51462 consensused 51400 bridges out of 51462 Done building consensus bridges Now we will determine through an iterative process what contigs of the assembly are present only once in the final genome Out of 51462 supposed single-copy contigs, 11975 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 39487 supposed single-copy contigs, 4719 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 34768 supposed single-copy contigs, 2333 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 32435 supposed single-copy contigs, 1244 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 31191 supposed single-copy contigs, 744 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 30447 supposed single-copy contigs, 412 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 30035 supposed single-copy contigs, 253 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 29782 supposed single-copy contigs, 148 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 29634 supposed single-copy contigs, 92 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 29542 supposed single-copy contigs, 59 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 29483 supposed single-copy contigs, 37 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 29446 supposed single-copy contigs, 30 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 29416 supposed single-copy contigs, 21 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 29395 supposed single-copy contigs, 13 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 29382 supposed single-copy contigs, 8 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 29374 supposed single-copy contigs, 6 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 29368 supposed single-copy contigs, 4 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 29364 supposed single-copy contigs, 3 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 29361 supposed single-copy contigs, 3 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 29358 supposed single-copy contigs, 3 were not actually haploid. Recomputing until all the single-copy contigs are robust. Out of 29355 supposed single-copy contigs, 1 were not actually haploid. Recomputing until all the single-copy contigs are robust. Let's move on to actually untangling the graph Processed 0 contigs out of 102461 , while untangling with long reads Processed 1000 contigs out of 102461 , while untangling with long reads Processed 2000 contigs out of 102461 , while untangling with long reads Processed 3000 contigs out of 102461 , while untangling with long reads Processed 4000 contigs out of 102461 , while untangling with long reads Debug WARNING, ['271376', '16521597', '272120', '77390718', '7591698'] : looking for ['7591698'] 0 from ['77390718'] among [['7591698'], ['14360169']] [1, 1] 271376 <16521597>272120<77390718>7591698 Processed 5000 contigs out of 102461 , while untangling with long reads Processed 6000 contigs out of 102461 , while untangling with long reads Processed 7000 contigs out of 102461 , while untangling with long reads Processed 8000 contigs out of 102461 , while untangling with long reads Processed 9000 contigs out of 102461 , while untangling with long reads Processed 10000 contigs out of 102461 , while untangling with long reads Processed 11000 contigs out of 102461 , while untangling with long reads Processed 12000 contigs out of 102461 , while untangling with long reads Processed 13000 contigs out of 102461 , while untangling with long reads Processed 14000 contigs out of 102461 , while untangling with long reads Processed 15000 contigs out of 102461 , while untangling with long reads Debug WARNING, ['4229900', '16324833', '16324841', '16324849', '16324859', '16324867', '16324873', '16324879', '16324885', '16324897', '16324903', '16472453', '16445741', '16445753', '16445759', '16445765', '16538773', '16538779', '16538787', '16538797', '16538803', '16538809', '81112111', '16538809', '53260245', '82122851', '6222572', '4773173'] : looking for ['81112111'] 0 from ['16538809'] among [['81112111'], ['16538817']] [1, 0] 4229900 >16324833>16324841>16324849>16324859>16324867>16324873>16324879>16324885>16324897>16324903>16472453>16445741>16445753>16445759>16445765>16538773>16538779>16538787>16538797>16538803>16538809>81112111<16538809<53260245<82122851<6222572>4773173 Processed 16000 contigs out of 102461 , while untangling with long reads Processed 17000 contigs out of 102461 , while untangling with long reads Processed 18000 contigs out of 102461 , while untangling with long reads Processed 19000 contigs out of 102461 , while untangling with long reads Processed 20000 contigs out of 102461 , while untangling with long reads Processed 21000 contigs out of 102461 , while untangling with long reads Processed 22000 contigs out of 102461 , while untangling with long reads Processed 23000 contigs out of 102461 , while untangling with long reads Processed 24000 contigs out of 102461 , while untangling with long reads Processed 25000 contigs out of 102461 , while untangling with long reads Processed 26000 contigs out of 102461 , while untangling with long reads Traceback (most recent call last): File "/Users/jff/.local/bin/graphunzip.py", line 335, in main() File "/Users/jff/.local/bin/graphunzip.py", line 312, in main bridge_with_long_reads(segments, names, cn, lrFile, supported_links2, refHaploidy, multiplicities) File "/Users/jff/.local/src/GraphUnzip/solve_with_long_reads.py", line 120, in bridge_with_long_reads unzip_graph_with_bridges(segments, non_overlapping_bridges, copiesnumber, haploidContigs, haploidContigsNames, names, supported_links2, minimum_supported_links, multiplicities, longContigs) File "/Users/jff/.local/src/GraphUnzip/solve_with_long_reads.py", line 436, in unzip_graph_with_bridges CIGAR = segments[names[contigs[0]]].CIGARs[end][segment.find_this_link(segments[oldContigsIndices[1]], nextEnd, segments[names[contigs[0]]].links[end], segments[names[contigs[0]]].otherEndOfLinks[end])] IndexError: list index out of range

RolandFaure commented 2 years ago

Hi Jean-François, I found the origin of the problem: it was that SPAligner does not always output alignments compatible with the assembly graph. I added a check in GraphUnzip so that alignments incompatible with the graph are discarded with a warning message. You can now use the new version, thank you for bringing up this problem