Path to genome folder specified as: /gscratch/srlab/sam/data/C_gigas/genomes/ Aligner to be used: Bowtie 2 (default) Writing bisulfite genomes out into a single MFA (multi FastA) file Bismark Genome Preparation - Step I: Preparing folders Path to Bowtie 2 specified: /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/ Bisulfite Genome Indexer version v0.19.0 (last modified 07 November 2016) Created Bisulfite Genome folder /gscratch/srlab/sam/data/C_gigas/genomes/Bisulfite_Genome/ Created Bisulfite Genome folder /gscratch/srlab/sam/data/C_gigas/genomes/Bisulfite_Genome/CT_conversion/ Created Bisulfite Genome folder /gscratch/srlab/sam/data/C_gigas/genomes/Bisulfite_Genome/GA_conversion/ Step I - Prepare genome folders - completed Bismark Genome Preparation - Step II: Bisulfite converting reference genome conversions performed: chromosome C->T G->A scaffold22 302346 304918 scaffold1009 275878 273965 scaffold150 277786 280257 scaffold1024 260644 259035 scaffold102 260155 259856 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48 C14366 49 47 C14362 46 46 C14354 31 36 C14370 36 38 C14334 57 32 C14344 69 27 C14332 55 46 C14322 38 30 C14326 57 31 C14340 35 53 C14320 48 32 C14312 40 57 C14304 38 44 C14306 38 42 C14308 33 30 C14316 43 38 C14288 32 38 C14282 44 49 C14278 47 36 C14276 39 59 C14292 34 45 C14300 43 33 C14250 35 62 C14248 36 49 C14274 39 41 C14272 44 27 C14242 39 38 C14232 46 54 C14228 42 33 C14226 42 35 C14230 32 43 C14236 79 56 C14238 39 31 C14218 58 50 C14210 60 59 C14222 32 25 C14214 41 47 C14220 43 27 C14212 46 34 C14182 49 50 C14184 52 53 C14204 31 43 C14202 46 42 C14178 43 32 C14186 45 36 C14188 47 42 C14180 55 41 C14166 30 59 C14170 58 36 C14174 56 43 C14152 37 35 C14162 52 35 C14150 23 27 C14134 18 78 C14138 27 27 C14142 36 50 C14128 46 29 C14132 23 63 C14108 34 52 C14118 38 33 C14098 22 49 C14104 31 44 C14086 30 52 C14110 33 43 C14068 66 44 C14080 35 38 C14064 31 32 C14076 71 44 C14066 43 35 C14074 42 40 scaffold899 30 34 C14072 73 40 C14060 35 32 scaffold153 28 43 C14022 41 42 C14046 40 34 C14028 41 39 C14044 46 21 C14050 49 43 C14032 35 35 C14036 31 27 C14004 39 41 C14008 42 34 C14010 35 31 C14016 30 36 C13980 60 44 C13990 42 34 C13972 56 60 C13978 33 28 C13992 39 44 C13976 38 36 C13994 39 32 C13954 40 33 C13962 33 39 C13956 23 50 C13950 35 53 C13940 31 47 C13938 42 31 C13930 35 64 C13946 51 39 C13958 27 34 C13920 34 25 C13924 28 28 C13904 40 43 C13908 31 46 C13916 35 39 C13892 57 22 C13896 33 26 C13888 26 80 C13898 36 27 C13854 41 33 C13878 62 40 C13872 46 34 C13874 40 41 C13866 47 33 C13880 48 45 C13860 41 28 C13868 40 33 C13852 28 28 C13810 33 44 C13814 35 47 C13842 30 20 C13834 42 38 C13838 30 53 C13844 52 37 C13836 39 41 C13800 48 35 C13780 53 42 C13798 49 28 C13802 43 33 C13786 39 42 C13788 61 34 C13804 43 36 C13754 32 35 C13766 29 32 C13762 30 45 C13732 30 45 C13736 49 56 C13712 46 38 C13720 40 41 C13716 47 32 C13724 41 19 C13694 26 34 C13688 26 48 scaffold238 63 36 C13690 35 42 C13676 44 30 C13648 37 31 C13670 41 27 C13642 36 32 C13666 32 36 C13652 21 45 C13668 36 31 C13654 33 38 C13650 26 34 C13624 43 25 C13612 45 33 C13620 35 35 C13638 35 39 C13616 38 46 C13632 35 55 C13614 42 41 C13634 41 53 C13606 48 40 C13584 54 38 C13600 49 45 C13598 48 35 C13592 43 38 C13590 39 35 C13572 32 17 C13582 49 36 C13576 25 33 C13578 45 39 C13570 50 45 C13580 45 15 C13552 44 48 C13516 33 44 C13508 48 37 C13520 36 28 C13532 51 48 C13524 49 31 C13542 30 28 C13546 22 37 C13526 37 32 C13534 31 31 C13512 47 27 C13528 30 37 C13482 70 39 C13466 41 29 C13464 34 36 C13460 35 27 C13476 32 31 C13490 29 34 C13500 28 29 C13496 26 40 C13486 37 27 C13488 51 40 C13492 38 35 C13462 27 32 C13442 36 30 C13450 30 30 C13448 29 40 C13452 45 33 C13440 39 35 C13424 65 45 C13428 48 46 C13414 34 35 C13418 32 31 C13406 44 31 C13396 40 54 C13410 53 38 C13408 40 30 C13416 35 22 C13368 40 34 C13390 29 37 C13380 50 47 C13356 24 48 C13382 34 60 C13384 32 36 C13372 30 31 C13392 36 25 C13370 31 37 C13360 31 22 C13320 41 41 C13326 56 25 C13336 34 32 C13324 54 39 C13344 45 35 C13328 21 69 C13346 40 42 C13348 41 21 C13330 39 36 C13298 37 44 C13300 39 36 C13316 29 33 C13292 42 35 C13294 44 37 C13268 36 38 C13266 31 46 C13278 53 35 C13256 20 32 C13288 35 36 C13272 57 43 C13258 47 52 C13284 36 23 C13254 39 45 C13264 40 41 C13274 37 34 C13260 47 36 C13236 26 63 C13242 28 35 C13234 40 30 C13224 25 33 C13218 59 25 C13244 39 35 C13220 25 37 C13228 36 73 C13248 40 33 C13182 35 59 C13176 52 34 C13204 43 34 C13180 47 40 C13194 44 30 C13186 44 32 C13210 43 41 C13202 31 35 C13208 41 25 C13178 45 29 C13198 55 34 C13206 49 38 C13140 48 39 C13148 34 49 C13134 32 33 C13172 34 31 C13156 39 40 C13132 30 29 C13164 37 26 C13170 21 43 C13136 48 38 C13094 40 36 C13116 46 33 C13106 48 52 C13102 32 46 C13118 22 56 C13130 20 29 C13092 24 34 C13098 39 43 C13120 36 36 C13128 32 20 C13100 30 34 C13042 39 32 C13078 37 39 C13076 30 38 C13086 30 41 C13052 54 30 C13046 42 31 C13048 47 45 C13080 34 43 C13050 44 19 C13068 29 25 C13054 40 51 C13082 25 39 C13074 32 26 C13040 36 32 C13030 47 35 C13012 35 34 C13016 23 41 C13034 50 23 C13028 39 44 C13020 54 29 C13026 40 36 C12994 21 50 C12972 34 31 C12984 38 25 C12974 42 40 C12970 26 45 C12948 12 42 C12964 34 17 C12960 35 28 C12916 41 44 C12924 21 24 C12932 48 37 C12958 41 24 C12918 28 25 C12950 35 60 C12956 28 37 C12938 53 31 C12954 24 25 C12928 48 30 C12942 32 41 C12912 39 48 C12934 35 37 C12930 58 39 C12882 31 37 C12890 28 41 C12872 40 37 C12886 30 39 C12880 30 34 C12906 27 36 C12876 32 37 C12870 18 28 C12888 51 41 C12836 40 30 C12832 28 42 C12856 34 26 C12866 37 39 C12840 26 48 C12838 46 39 C12848 36 34 C12834 28 30 C12860 51 42 C12842 31 32 C12802 32 31 C12806 27 34 C12786 24 18 C12788 36 21 C12810 26 32 C12828 29 51 C12822 36 31 C12824 35 28 C12814 32 35 C12830 29 25 C12798 44 44 C12782 38 31 C12766 34 35 C12768 23 40 C12738 18 38 C12758 35 38 C12742 37 28 C12774 29 43 C12760 27 33 C12772 38 37 C12764 32 42 C12780 33 36 C12778 32 40 C12784 35 37 C12748 25 37 C12728 37 28 C12722 34 50 C12706 40 41 C12718 30 60 C12732 28 25 C12726 25 59 C12708 44 43 C12730 42 23 C12724 62 21 C12734 34 52 Total number of conversions performed: C->T: 82182542 G->A: 82217681 Step II - Genome bisulfite conversions - completed Bismark Genome Preparation - Step III: Launching the Bowtie 2 indexer Please be aware that this process can - depending on genome size - take several hours! Preparing indexing of CT converted genome in /gscratch/srlab/sam/data/C_gigas/genomes/Bisulfite_Genome/CT_conversion/ Parent process: Starting to index C->T converted genome with the following command: /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/bowtie2-build -f genome_mfa.CT_conversion.fa BS_CT Settings: Output files: "BS_CT.*.bt2" Line rate: 6 (line is 64 bytes) Lines per side: 1 (side is 64 bytes) Offset rate: 4 (one in 16) FTable chars: 10 Strings: unpacked Max bucket size: default Max bucket size, sqrt multiplier: default Max bucket size, len divisor: 4 Difference-cover sample period: 1024 Endianness: little Actual local endianness: little Sanity checking: disabled Assertions: disabled Random seed: 0 Sizeofs: void*:8, int:4, long:8, size_t:8 Input files DNA, FASTA: genome_mfa.CT_conversion.fa Building a SMALL index Reading reference sizes Preparing indexing of GA converted genome in /gscratch/srlab/sam/data/C_gigas/genomes/Bisulfite_Genome/GA_conversion/ Child process: Starting to index G->A converted genome with the following command: /gscratch/srlab/programs/bowtie2-2.3.4.1-linux-x86_64/bowtie2-build -f genome_mfa.GA_conversion.fa BS_GA Settings: Output files: "BS_GA.*.bt2" Line rate: 6 (line is 64 bytes) Lines per side: 1 (side is 64 bytes) Offset rate: 4 (one in 16) FTable chars: 10 Strings: unpacked Max bucket size: default Max bucket size, sqrt multiplier: default Max bucket size, len divisor: 4 Difference-cover sample period: 1024 Endianness: little Actual local endianness: little Sanity checking: disabled Assertions: disabled Random seed: 0 Sizeofs: void*:8, int:4, long:8, size_t:8 Input files DNA, FASTA: genome_mfa.GA_conversion.fa Building a SMALL index Reading reference sizes Time reading reference sizes: 00:00:04 Calculating joined length Writing header Reserving space for joined string Joining reference sequences Time reading reference sizes: 00:00:04 Calculating joined length Writing header Reserving space for joined string Joining reference sequences Time to join reference sequences: 00:00:02 bmax according to bmaxDivN setting: 122962645 Using parameters --bmax 92221984 --dcv 1024 Doing ahead-of-time memory usage test Passed! Constructing with these parameters: --bmax 92221984 --dcv 1024 Constructing suffix-array element generator Building DifferenceCoverSample Building sPrime Building sPrimeOrder V-Sorting samples Time to join reference sequences: 00:00:03 bmax according to bmaxDivN setting: 122962645 Using parameters --bmax 92221984 --dcv 1024 Doing ahead-of-time memory usage test Passed! Constructing with these parameters: --bmax 92221984 --dcv 1024 Constructing suffix-array element generator Building DifferenceCoverSample Building sPrime Building sPrimeOrder V-Sorting samples V-Sorting samples time: 00:00:09 Allocating rank array Ranking v-sort output V-Sorting samples time: 00:00:08 Allocating rank array Ranking v-sort output Ranking v-sort output time: 00:00:02 Invoking Larsson-Sadakane on ranks Ranking v-sort output time: 00:00:02 Invoking Larsson-Sadakane on ranks Invoking Larsson-Sadakane on ranks time: 00:00:04 Sanity-checking and returning Building samples Reserving space for 12 sample suffixes Generating random suffixes QSorting 12 sample offsets, eliminating duplicates QSorting sample offsets, eliminating duplicates time: 00:00:00 Multikey QSorting 12 samples (Using difference cover) Multikey QSorting samples time: 00:00:00 Calculating bucket sizes Splitting and merging Splitting and merging time: 00:00:00 Avg bucket size: 4.91851e+08 (target: 92221983) Converting suffix-array elements to index image Allocating ftab, absorbFtab Entering Ebwt loop Getting block 1 of 1 No samples; assembling all-inclusive block Sorting block of length 491850583 for bucket 1 (Using difference cover) Invoking Larsson-Sadakane on ranks time: 00:00:04 Sanity-checking and returning Building samples Reserving space for 12 sample suffixes Generating random suffixes QSorting 12 sample offsets, eliminating duplicates QSorting sample offsets, eliminating duplicates time: 00:00:00 Multikey QSorting 12 samples (Using difference cover) Multikey QSorting samples time: 00:00:00 Calculating bucket sizes Splitting and merging Splitting and merging time: 00:00:00 Avg bucket size: 4.91851e+08 (target: 92221983) Converting suffix-array elements to index image Allocating ftab, absorbFtab Entering Ebwt loop Getting block 1 of 1 No samples; assembling all-inclusive block Sorting block of length 491850583 for bucket 1 (Using difference cover) Sorting block time: 00:02:25 Returning block of 491850584 for bucket 1 Sorting block time: 00:02:27 Returning block of 491850584 for bucket 1 Exited Ebwt loop fchr[A]: 0 fchr[C]: 245953980 fchr[G]: 328136522 fchr[T]: 328136522 fchr[$]: 491850583 Exiting Ebwt::buildToDisk() Returning from initFromVector Wrote 168716543 bytes to primary EBWT file: BS_GA.1.bt2 Wrote 122962652 bytes to secondary EBWT file: BS_GA.2.bt2 Re-opening _in1 and _in2 as input streams Returning from Ebwt constructor Headers: len: 491850583 bwtLen: 491850584 sz: 122962646 bwtSz: 122962646 lineRate: 6 offRate: 4 offMask: 0xfffffff0 ftabChars: 10 eftabLen: 20 eftabSz: 80 ftabLen: 1048577 ftabSz: 4194308 offsLen: 30740662 offsSz: 122962648 lineSz: 64 sideSz: 64 sideBwtSz: 48 sideBwtLen: 192 numSides: 2561722 numLines: 2561722 ebwtTotLen: 163950208 ebwtTotSz: 163950208 color: 0 reverse: 0 Total time for call to driver() for forward index: 00:03:16 Reading reference sizes Exited Ebwt loop fchr[A]: 0 fchr[C]: 163736299 fchr[G]: 163736299 fchr[T]: 245953980 fchr[$]: 491850583 Exiting Ebwt::buildToDisk() Returning from initFromVector Wrote 168716543 bytes to primary EBWT file: BS_CT.1.bt2 Wrote 122962652 bytes to secondary EBWT file: BS_CT.2.bt2 Re-opening _in1 and _in2 as input streams Returning from Ebwt constructor Headers: len: 491850583 bwtLen: 491850584 sz: 122962646 bwtSz: 122962646 lineRate: 6 offRate: 4 offMask: 0xfffffff0 ftabChars: 10 eftabLen: 20 eftabSz: 80 ftabLen: 1048577 ftabSz: 4194308 offsLen: 30740662 offsSz: 122962648 lineSz: 64 sideSz: 64 sideBwtSz: 48 sideBwtLen: 192 numSides: 2561722 numLines: 2561722 ebwtTotLen: 163950208 ebwtTotSz: 163950208 color: 0 reverse: 0 Total time for call to driver() for forward index: 00:03:20 Reading reference sizes Time reading reference sizes: 00:00:03 Calculating joined length Writing header Reserving space for joined string Joining reference sequences Time reading reference sizes: 00:00:04 Calculating joined length Writing header Reserving space for joined string Joining reference sequences Time to join reference sequences: 00:00:08 Time to reverse reference sequence: 00:00:00 bmax according to bmaxDivN setting: 122962645 Using parameters --bmax 92221984 --dcv 1024 Doing ahead-of-time memory usage test Passed! Constructing with these parameters: --bmax 92221984 --dcv 1024 Constructing suffix-array element generator Building DifferenceCoverSample Building sPrime Building sPrimeOrder V-Sorting samples Time to join reference sequences: 00:00:09 Time to reverse reference sequence: 00:00:01 bmax according to bmaxDivN setting: 122962645 Using parameters --bmax 92221984 --dcv 1024 Doing ahead-of-time memory usage test Passed! Constructing with these parameters: --bmax 92221984 --dcv 1024 Constructing suffix-array element generator Building DifferenceCoverSample Building sPrime Building sPrimeOrder V-Sorting samples V-Sorting samples time: 00:00:08 Allocating rank array Ranking v-sort output Ranking v-sort output time: 00:00:02 Invoking Larsson-Sadakane on ranks Invoking Larsson-Sadakane on ranks time: 00:00:03 Sanity-checking and returning Building samples Reserving space for 12 sample suffixes Generating random suffixes QSorting 12 sample offsets, eliminating duplicates QSorting sample offsets, eliminating duplicates time: 00:00:00 Multikey QSorting 12 samples (Using difference cover) Multikey QSorting samples time: 00:00:00 Calculating bucket sizes Splitting and merging Splitting and merging time: 00:00:00 Avg bucket size: 4.91851e+08 (target: 92221983) Converting suffix-array elements to index image Allocating ftab, absorbFtab Entering Ebwt loop Getting block 1 of 1 No samples; assembling all-inclusive block V-Sorting samples time: 00:00:09 Allocating rank array Ranking v-sort output Sorting block of length 491850583 for bucket 1 (Using difference cover) Ranking v-sort output time: 00:00:02 Invoking Larsson-Sadakane on ranks Invoking Larsson-Sadakane on ranks time: 00:00:03 Sanity-checking and returning Building samples Reserving space for 12 sample suffixes Generating random suffixes QSorting 12 sample offsets, eliminating duplicates QSorting sample offsets, eliminating duplicates time: 00:00:00 Multikey QSorting 12 samples (Using difference cover) Multikey QSorting samples time: 00:00:00 Calculating bucket sizes Splitting and merging Splitting and merging time: 00:00:00 Avg bucket size: 4.91851e+08 (target: 92221983) Converting suffix-array elements to index image Allocating ftab, absorbFtab Entering Ebwt loop Getting block 1 of 1 No samples; assembling all-inclusive block Sorting block of length 491850583 for bucket 1 (Using difference cover) Sorting block time: 00:02:25 Returning block of 491850584 for bucket 1 Sorting block time: 00:02:26 Returning block of 491850584 for bucket 1 Exited Ebwt loop fchr[A]: 0 fchr[C]: 245953980 fchr[G]: 328136522 fchr[T]: 328136522 fchr[$]: 491850583 Exiting Ebwt::buildToDisk() Returning from initFromVector Wrote 168716543 bytes to primary EBWT file: BS_GA.rev.1.bt2 Wrote 122962652 bytes to secondary EBWT file: BS_GA.rev.2.bt2 Re-opening _in1 and _in2 as input streams Returning from Ebwt constructor Headers: len: 491850583 bwtLen: 491850584 sz: 122962646 bwtSz: 122962646 lineRate: 6 offRate: 4 offMask: 0xfffffff0 ftabChars: 10 eftabLen: 20 eftabSz: 80 ftabLen: 1048577 ftabSz: 4194308 offsLen: 30740662 offsSz: 122962648 lineSz: 64 sideSz: 64 sideBwtSz: 48 sideBwtLen: 192 numSides: 2561722 numLines: 2561722 ebwtTotLen: 163950208 ebwtTotSz: 163950208 color: 0 reverse: 1 Total time for backward call to driver() for mirror index: 00:03:19 Exited Ebwt loop fchr[A]: 0 fchr[C]: 163736299 fchr[G]: 163736299 fchr[T]: 245953980 fchr[$]: 491850583 Exiting Ebwt::buildToDisk() Returning from initFromVector Wrote 168716543 bytes to primary EBWT file: BS_CT.rev.1.bt2 Wrote 122962652 bytes to secondary EBWT file: BS_CT.rev.2.bt2 Re-opening _in1 and _in2 as input streams Returning from Ebwt constructor Headers: len: 491850583 bwtLen: 491850584 sz: 122962646 bwtSz: 122962646 lineRate: 6 offRate: 4 offMask: 0xfffffff0 ftabChars: 10 eftabLen: 20 eftabSz: 80 ftabLen: 1048577 ftabSz: 4194308 offsLen: 30740662 offsSz: 122962648 lineSz: 64 sideSz: 64 sideBwtSz: 48 sideBwtLen: 192 numSides: 2561722 numLines: 2561722 ebwtTotLen: 163950208 ebwtTotSz: 163950208 color: 0 reverse: 1 Total time for backward call to driver() for mirror index: 00:03:31