archive:computer_resources:assemblies
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archive:computer_resources:assemblies [2011/05/31 20:28] karplus [slug/] More on mitochondrial genome |
archive:computer_resources:assemblies [2015/09/02 16:43] ceisenhart ↷ Page moved from computer_resources:assemblies to archive:computer_resources:assemblies |
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| * newbler-assembly2/ is a second de novo assembly using Newbler, starting from the cleaned reads of newbler-clean1/sff_cleaned/no_Hyp.sff It gets 42 contigs and 2,449,409 bases. | * newbler-assembly2/ is a second de novo assembly using Newbler, starting from the cleaned reads of newbler-clean1/sff_cleaned/no_Hyp.sff It gets 42 contigs and 2,449,409 bases. | ||
| * newbler-assembly3/ starts from the same sff file as newbler-assembly2/ but raises the expected coverage to 60 (close to actual coverage). It gets 41 contigs and 2,449,426 bases, still more than the old version of Newbler got after similar cleaning. The contigs have been mapped to the finished genome (using megablast, blastn, blat, and pluck-scripts/find-dna-differences). All the contigs map cleanly to the finished genome. If contigs map to more than one place, find-dna-differences may (incorrectly) report it as not mapping. | * newbler-assembly3/ starts from the same sff file as newbler-assembly2/ but raises the expected coverage to 60 (close to actual coverage). It gets 41 contigs and 2,449,426 bases, still more than the old version of Newbler got after similar cleaning. The contigs have been mapped to the finished genome (using megablast, blastn, blat, and pluck-scripts/find-dna-differences). All the contigs map cleanly to the finished genome. If contigs map to more than one place, find-dna-differences may (incorrectly) report it as not mapping. | ||
| - | * map-colorspace3/ uses the [[bioinformatic_tools:pluck-scripts|pluck-scripts]] script map-colorspace to map the SOLiD mate-pair reads onto the contigs of the newbler-assembly3/ run. The intent is to find what contigs join to what other ones. The numbering starts with 3, not 1, so that the map-colorspace directories correspond to the newbler-assembly directories that they are mapping onto. | + | * map-colorspace3/ uses the [[archive:bioinformatic_tools:pluck-scripts|pluck-scripts]] script map-colorspace to map the SOLiD mate-pair reads onto the contigs of the newbler-assembly3/ run. The intent is to find what contigs join to what other ones. The numbering starts with 3, not 1, so that the map-colorspace directories correspond to the newbler-assembly directories that they are mapping onto. |
| * newbler-partial3/ assembled the partially-assembled reads of newbler-assembly3/ to see if any extended or connected contigs. Seven of the 131 new contigs could be used to extend newbler-assembly3/ contigs, but none spanned 2 contigs. | * newbler-partial3/ assembled the partially-assembled reads of newbler-assembly3/ to see if any extended or connected contigs. Seven of the 131 new contigs could be used to extend newbler-assembly3/ contigs, but none spanned 2 contigs. | ||
| * newbler-assembly4/ starts from the same sff file as newbler-assembly2/ and newbler-assembly3/ but adds the contigs of newbler-partial3/ as extra reads. This did not help, getting 45 contigs and 2,449,287 bases. | * newbler-assembly4/ starts from the same sff file as newbler-assembly2/ and newbler-assembly3/ but adds the contigs of newbler-partial3/ as extra reads. This did not help, getting 45 contigs and 2,449,287 bases. | ||
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| * so ran with filling -R to get 12k maxcontig. | * so ran with filling -R to get 12k maxcontig. | ||
| * Then ran the scaffolding steps with 200bp insert size. | * Then ran the scaffolding steps with 200bp insert size. | ||
| - | * For all steps, used low default cutoffs since our 10x coverage | + | * For all steps, used low default cutoffs since our 10x coverage is not high. 21k max scaffold size. |
| - | * is not high. 21k max scaffold size. Estimated | + | * Estimated genome size is around 3G. |
| - | * genome size is around 3G. The 4 steps are | + | * The 4 steps are |
| - pregraph (3.5 to 4.5 hours for 30 to 60 cpus) | - pregraph (3.5 to 4.5 hours for 30 to 60 cpus) | ||
| - contig (1.3 hours) | - contig (1.3 hours) | ||
| - map (0.6 hours with 60cpus) - paired ends | - map (0.6 hours with 60cpus) - paired ends | ||
| - scaff (1 hour with 60cpus) | - scaff (1 hour with 60cpus) | ||
| - | * barcode-of-life/ attempt to assemble the mitochondrial genome, with particular emphasis on the gene for mitochondrial cytochrome c oxidase subunit I protein I (CO1), which is used for the "barcode of life". [[http://www.boldsystems.org/|BOLD (barcode of life database)]] | + | * barcode-of-life/ attempt to assemble the mitochondrial genome, documented on its own page: [[computer_resources:assemblies:mitochondrion]] |
| - | * Started with a search of SOAPdenovo-assembly1/k31/soapSlug.scafSeq for scaffolds that matched examples from other mollusks. | + | * SOAPdenovo-assembly2/ Assembly with new + old Illumina and 454 data. |
| - | * Looked for 454 reads that extended or joined contigs in scaffold | + | * SOAPdenovo 1.05 - can handle gzipped fastq files. |
| - | * Repeated (sometimes using more sensitive searches) until no more credible scaffolds from the SOAPdenovo-assembly1/k31/ assembly nor 454 reads were found. | + | * Runs with k27, 31, 47, and 63 so far. 47 was the best overall. 63 got the longest contig (~14.9kb). |
| - | * The 454 coverage of the mitochondrion is so slight as to be nearly useless, so instead we can iterate: | + | * Run parameters: |
| - | - find all Illumina reads that map to the mitochondrial draft, using BWA | + | - pregraph: |
| - | - assemble them using SOAPdenovo. | + | * lowest count size of 2 (-d 2) |
| - | * It looks like the Illumina reads have about 228x coverage of the mitochondrion, but coverage is patchy, and it seems to be difficult to close the circle (at least with SOAPdenovo). | + | - contig: |
| - | * I have an almost complete mitochondrial genome, and I'm hoping that a few more iterations or some tricky assembly will close it into a clean circular genome. | + | * solve tiny repeats on (-R) |
| + | - map: | ||
| + | * all default | ||
| + | - scaff: | ||
| + | * intra-scaffold gap closure on (-F) | ||
| + | * Statistics for each kmer size assembly (using illumina and 454 data, using both for contig and scaffolding): | ||
| + | * k31: | ||
| + | * 1,298,372 scaffolds from 4,814,226 contigs sum up 632,702,276bp, with average length 487, 0 gaps filled | ||
| + | * 3,611,844 scaffolds&singleton sum up 1,133,413,022bp, with average length 313 | ||
| + | * the longest is 10,340bp,scaffold N50 is 442 bp, scaffold N90 is 148 bp | ||
| + | * k47: | ||
| + | * 871,819 scaffolds from 5,306,463 contigs sum up 530,762,874bp, with average length 608, 0 gaps filled | ||
| + | * 4,203,195 scaffolds&singleton sum up 1,296,678,043bp, with average length 308 | ||
| + | * the longest is 14,750bp,scaffold N50 is 458 bp, scaffold N90 is 140 bp | ||
| + | * k63: | ||
| + | * 270,887 scaffolds from 4,022,505 contigs sum up 139,720,415bp, with average length 515, 0 gaps filled | ||
| + | * 3,710,532 scaffolds&singleton sum up 690,332,560bp, with average length 186 | ||
| + | * the longest is 14,897bp,scaffold N50 is 232 bp, scaffold N90 is 112 bp | ||
archive/computer_resources/assemblies.txt · Last modified: 2015/09/02 16:53 by 92.247.181.31
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