Brugia malayi: Difference between revisions
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Revision as of 14:19, 15 February 2010
Articles
Genome Info
- 6 chromosomes: 1-5, XY ; diploit genome ~ 110M bp
- 30% GC,
- 32% coding, 15% repeats
Other sequences
- mitochondrion finished: 13,657 bp; 24% GC
- Wolbachia endosymbiont strain TRS from Brugia malayi strain wMel complete: 1,080,084 bp; 34%GC (New England Biolabs)
- Wolbachia endosymbiont strain wMel progress (TIGR)
- Rodent: some trace contamination; Example: Mus musculus is ~44%GC
contaminants min q1 q2 q3 max mean n50 sum 2929 200 527 675 820 8994 740.04 762 2167588
Genome Project
Brugia malayi has a diploid genome of approximately 110 Mb, organized in 6 pairs of chromosomes (five pairs of autosomes and one pair of sex chromosomes). In addition to the nuclear genome, B. malayi has a mitochondrial genome of about 14kb, and the genome of the harbored bacterial endosymbiont Wolbachia sp (1-2Mb).
The B. malayi genome project has been completed by The Institute for Genomic Research. Whole Genome Shotgun sequencing was used to obtain more than eight-fold coverage of the genome. The complete genome was assembled into approximately 8200 scaffolds and deposited in GenBank. The accession for the WGS project is AAQA00000000 and consists of sequences AAQA01000001-AAQA01029808. File location:
/fs/szasmg3/dpuiu/Brugia_malayi/Data/Bm.fasta ctgs min q1 q2 q3 max mean n50 sum 26879 200 836 1005 1495 611244 3241.17 18986 87119350
- TIGR Genome project (TRS strain)
Data
Original Traces
- 1.26M Sanger reads & 15 Libraries:
- NCBI TA
- NCBI TA FTP
SEQ_LIB_ID INSERT_SIZE INSERT_STDEV TRACE_TYPE_CODE 1047113828118 1000 300 WGS 13500 1047113856575 1000 300 PRIMERWALK 325 1047111632737 1258 377 PRIMERWALK 3 1047111632737 1258 377 WGS 305,906 1047111540304 1415 424 WGS 51772 1047112577106 1415 424 WGS 337,789 1047111718946 3123 936 WGS 47597 1047113358719 3123 936 PRIMERWALK 173 1047113358719 3123 936 WGS 246,185 1047174912885 3123 936 TRANSPOSON 1437 1047113570927 6000 1800 WGS 3193 1047111814561 7158 2147 WGS 219,306 1047111480027 17168 5150 WGS 4087 1047111488095 17168 5150 WGS 3434 1047111495007 17168 5150 WGS 3716 1047111501919 17168 5150 WGS 3697 1047111480605 22419 6725 WGS 4638 1047111516154 22419 6725 WGS 4004 1047111523212 22419 6725 WGS 3766 1047111530126 22419 6725 WGS 5686 1047113855421 23000 6900 WGS 1 total 1,260,215
FRG file:
- FRG.src : TI's
- FRG.acc: 2 ..
- DST.acc: 1260217, ... , 1260234
- Location
/fs/szasmg3/dpuiu/Brugia_malayi/Data/nucmer_seq/Bm-all.frg DST 15 FRG 1178192 LKG 530930 seqs min q1 q2 q3 max mean n50 sum 1,178,192 65 645 771 850 1214 724 800 853,847,771 => 8X
Problems:
- All library insert sizes are underestimated ???
- The contaminant reads align at ~91-93% id to the contaminant ctgs while the Mt/We reads align at 99% id to Mt/We finished seq. What %id thold to use for contaminant?
BACS
8,000 BAC clones @Children's Hospital Oakland Research Institute. (!!! no NCBI TA submission)
PITT FTP data
- 3.21M 454 reads
- 454 Titanium paired-end (NEW)
- 500Mbp per run in reads of ~350bp each.
- reads: 80bp to 500bp
- 30% - 50% of the reads contain two mate tags per read
- Each mate tag is ~150bp and the tags are separated by the 42bp "recombi" XLR linker sequence
- lib mean,std =~ 28K,6K
- duplication problem: 2-4 identical copies of each pair???
CBCB Location:
/fs/szattic-asmg4/brugia_malayi/Data/
FTP access:
lftp -u bma 136.142.191.201 pass: 6279 user: bma # empty as of --Dpuiu 12:04, 8 January 2010 (EST)
Elodie's table: /scratch1/brugia_malayi/brugia-sequencing-summary.txt.csv
# elodie's date protocol platform type description run_name Reads Mates 1 01/17/2008 WGS Standard Full run (2/2) Mix of worms (calibration of the machine) R_2008_01_31_18_01_35_FLX10070260_adminrig_ghedintestsample ? 0 2 07/01/2008 3Kb Standard Full single worm (pUC contamination) R_2008_08_06_13_52_29_FLX10070260_adminrig_080608_Ghedin-BrugiaLTPE1 492575 84341 3 09/11/2008 3kb Standard 4/8 wells single worm (pUC contamination) R_2008_09_19_14_17_55_FLX10070260_adminrig_091908_HATFULL-MIDrepeat_GHEDIN-LTPE1 263421 49258 4 10/01/2008 3Kb Standard Full Mix of worms (still pUC contamination) R_2008_10_14_15_06_50_FLX10070260_adminrig_101408_GHEDIN-Brugia-pool_LTPEtest 59711 5096 5 02/01/2009 WGS Standard 1/4 wells Mix of worms; regions 2 & 3 were myxoma R_2009_02_27_16_11_34_FLX10070260_adminrig_022709_GHEDIN ? 0 6 04/06/2009 WGS Standard 1/4 wells Mix of worms; with comp. bio run R_2009_04_15_14_46_56_FLX10070260_adminrig_041509_GHEDIN_r1-WGS1_r2-LMW4_r3-pool2compbio_r4-pool3compbio ? 0 7 05/01/2009 20Kb Titanium 7/8 wells Mix of worms R_2009_06_05_16_18_41_FLX10070260_adminrig_060509_GHEDIN_Brugia-gDNA-TI20kb1 631287 213524 8 10/28/2009 20Kb Titanium Full Mix of worms R_2009_10_22_15_30_12_FLX10070260_adminrig_102209_GHEDIN_Brugia20kb2 1095713 377547 9 Pending 3 Kb Titanium Full Mix of worms ? ? ? . Total 2542707 729766
SFF files counts:
run sffReads linker
1 R_2008_01_31_18_01_35_FLX10070260_adminrig_ghedintestsample/D_2008_01_31_18_01_35_FLX10070260_adminrig_FullAnalysis/sff/E4RA0X101.sff 272923
1 R_2008_01_31_18_01_35_FLX10070260_adminrig_ghedintestsample/D_2008_01_31_18_01_35_FLX10070260_adminrig_FullAnalysis/sff/E4RA0X102.sff 261899
2 R_2008_08_06_13_52_29_FLX10070260_adminrig_080608_Ghedin-BrugiaLTPE1/D_2009_02_12_22_12_04_j_SignalProcessing/sff/FEZH5RS01.sff 228204 flx
2 R_2008_08_06_13_52_29_FLX10070260_adminrig_080608_Ghedin-BrugiaLTPE1/D_2009_02_12_22_12_04_j_SignalProcessing/sff/FEZH5RS02.sff 264371 flx
3 R_2008_09_19_14_17_55_FLX10070260_adminrig_091908_HATFULL-MIDrepeat_GHEDIN-LTPE1/FHAVB5T02.sff 86862 flx
3 R_2008_09_19_14_17_55_FLX10070260_adminrig_091908_HATFULL-MIDrepeat_GHEDIN-LTPE1/FHAVB5T03.sff 87488 flx
3 R_2008_09_19_14_17_55_FLX10070260_adminrig_091908_HATFULL-MIDrepeat_GHEDIN-LTPE1/FHAVB5T04.sff 89071 flx
4 R_2008_10_14_15_06_50_FLX10070260_adminrig_101408_GHEDIN-Brugia-pool_LTPEtest/FIOXLOM01.sff 13695 flx
4 R_2008_10_14_15_06_50_FLX10070260_adminrig_101408_GHEDIN-Brugia-pool_LTPEtest/FIOXLOM02.sff 14197 flx
4 R_2008_10_14_15_06_50_FLX10070260_adminrig_101408_GHEDIN-Brugia-pool_LTPEtest/FIOXLOM03.sff 15515 flx
4 R_2008_10_14_15_06_50_FLX10070260_adminrig_101408_GHEDIN-Brugia-pool_LTPEtest/FIOXLOM04.sff 16304 flx
5 R_2009_02_27_16_11_34_FLX10070260_adminrig_022709_GHEDIN/FRLDXKV01.sff 18025
6 R_2009_04_15_14_46_56_FLX10070260_adminrig_041509_GHEDIN_r1-WGS1_r2-LMW4_r3-pool2compbio_r4-pool3compbio/D_2009_04_16_14_19_21_morty_fullProcessing/FT9KOI001.sff 118490
7 R_2009_06_05_16_18_41_FLX10070260_adminrig_060509_GHEDIN_Brugia-gDNA-TI20kb1/D_2009_06_08_15_32_36_compute-0-2_fullProcessing/sff/FW1OXFY01.sff 73807 tit
7 R_2009_06_05_16_18_41_FLX10070260_adminrig_060509_GHEDIN_Brugia-gDNA-TI20kb1/D_2009_06_08_15_32_36_compute-0-2_fullProcessing/sff/FW1OXFY02.sff 91698 tit
7 R_2009_06_05_16_18_41_FLX10070260_adminrig_060509_GHEDIN_Brugia-gDNA-TI20kb1/D_2009_06_08_15_32_36_compute-0-2_fullProcessing/sff/FW1OXFY03.sff 93878 tit
7 R_2009_06_05_16_18_41_FLX10070260_adminrig_060509_GHEDIN_Brugia-gDNA-TI20kb1/D_2009_06_08_15_32_36_compute-0-2_fullProcessing/sff/FW1OXFY04.sff 90232 tit
7 R_2009_06_05_16_18_41_FLX10070260_adminrig_060509_GHEDIN_Brugia-gDNA-TI20kb1/D_2009_06_08_15_32_36_compute-0-2_fullProcessing/sff/FW1OXFY05.sff 97065 tit
7 R_2009_06_05_16_18_41_FLX10070260_adminrig_060509_GHEDIN_Brugia-gDNA-TI20kb1/D_2009_06_08_15_32_36_compute-0-2_fullProcessing/sff/FW1OXFY06.sff 94326 tit
7 R_2009_06_05_16_18_41_FLX10070260_adminrig_060509_GHEDIN_Brugia-gDNA-TI20kb1/D_2009_06_08_15_32_36_compute-0-2_fullProcessing/sff/FW1OXFY07.sff 90281 tit
8 R_2009_10_22_15_30_12_FLX10070260_adminrig_102209_GHEDIN_Brugia20kb2/F4H5CMB01.sff 551263 tit
8 R_2009_10_22_15_30_12_FLX10070260_adminrig_102209_GHEDIN_Brugia20kb2/F4H5CMB02.sff 544450 tit
total 3214044
- Libraries
lib mea std reads mates 1 E4RA0X101 0 0 271066 0 1 E4RA0X102 0 0 260166 0 2 FEZH5RS01 3000 300 181035 18676 2 FEZH5RS02 3000 300 211064 22270 3 FHAVB5T02 3000 300 68708 7850 3 FHAVB5T03 3000 300 69306 8227 4 FHAVB5T04 3000 300 70028 8353 4 FIOXLOM01 3000 300 10921 0 4 FIOXLOM02 3000 300 11157 0 4 FIOXLOM03 3000 300 12197 0 4 FIOXLOM04 3000 300 12727 0 5 FRLDXKV01 0 0 17349 0 6 FT9KOI001 0 0 108826 0 7 FW1OXFY01 20000 2000 86127 15825 7 FW1OXFY02 20000 2000 106911 19668 7 FW1OXFY03 20000 2000 109874 20396 7 FW1OXFY04 20000 2000 104797 18933 7 FW1OXFY05 20000 2000 113716 20649 7 FW1OXFY06 20000 2000 110693 20326 7 FW1OXFY07 20000 2000 105931 19176 8 F4H5CMB01 20000 2000 626046 109918 8 F4H5CMB02 20000 2000 628432 118903 total . . 3297077 429170
Location:
/fs/szattic-asmg4/brugia_malayi/Data/Frg/
#clr of the seqs in the frg files
seqs min q1 q2 q3 max mean n50 sum
all 3,297,077 3 156 248 301 2043 244 275 806,091,347 => 8X
mated 858,340 64 107 156 223 612 171 201 147,070,298
unmated 2,438,737 2 207 261 335 2042 268 286 655,723,972
Contaminant search
Sanger 454 jird 31,501 197,420 Mt 1,507 2,634 We 49,014 23,249
Assemblies
TIGR
- 9X coverage, 856K Sanger traces => 8,200 scaff & 29,808 ctg (avg. scaff=~10K & avg ctg=~3K)
- "scaffolds totaling ~71 Mb of data with a further ~17.5 Mb of contigs not integrated into any scaffold (orphan contigs)" (Science 2007)
- NCBI AAQA00000000 AAQA01000001-AAQA01029808
* 26,879 good ctgs * 2,929 jird contaminants (Example: AAQA01001321 : mouse 99%id hits)
good ctg len
#elem min max mean median n50 sum
all 26879 200 611244 3241 1005 19005 87119350
10K+ 1224 10036 611244 41018 23135 60727 50206329
good ctg GC%
#elem min max mean median n50
all 26878 0.00 72.30 28.86 28.56 29.46
10K+ 1224 24.38 38.44 30.38 30.43 30.62
contaminant ctg len
#elem min max mean median n50 sum
all 2929 200 8994 740 675 763 2167588
contaminant ctg GC%
#elem min max mean median n50
all 2929 18.09 75.96 44.1 43.59 44.80
CBCB
- test wgs 5.1 on filtered Sanger reads
- better assembly than the published one
- Location:
/fs/szasmg3/dpuiu/Brugia_malayi/Assembly/Bm/2008_0826_CA
#elem min max mean median n50 sum ctg 12753 273 376744 6113 1632 24765 77964006 ctg(10K+) 1553 10015 376744 33868 23216 40618 52597039 deg 9661 65 72494 1241 949 1008 11988997 ctg+deg 22414 65 376744 4013 1201 19411 89953003 ctg+deg(10K+) 1651 10015 376744 33019 22612 40060 54514638 scf 10317 935 3890532 8019 1538 41772 82730474 scf2K+ 3656 2001 3890532 20189 5733 50293 73813083 reads 1178192(100%) singletons 134119 (11.43%)
PITT
- Best so far
- Date: 11/05/08
- Location:
/fs/szasmg3/dpuiu/Brugia_malayi/Assembly/PITT/brugia_assemblies_110508.fasta
- Stats:
elem min q1 q2 q3 max mean n50 sum scf 3170 2000 2917 4483 14471 6534162 22916 112914 72643770
Files
/fs/szattic/asmg1/adelcher/Genomes/Brugia : Art's files /fs/sztmpscratch/cole/tarchive_download/brugia_malay : Cole's files /fs/szasmg3/dpuiu/Brugia_malayi/ : Daniela's files /scratch1/brugia_malayi/Data/ : ftp PITT data /fs/szattic-asmg4/brugia_malayi : ftp PITT data (as well)