Helicobacter pylori

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Revision as of 15:04, 21 October 2009 by Dpuiu (talk | contribs) (→‎Wustl)
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Data

Wustl

NCBI complete genomes

  • Genome info
       id             len     gc%
    1  NC_000915.1    1667867 38.87  Helicobacter pylori 26695
    2  NC_000921.1    1643831 39.19  Helicobacter pylori J99
    3  NC_008086.1    1596366 39.08  Helicobacter pylori HPAG1
    4  NC_010698.2    1608548 38.91  Helicobacter pylori Shi470
    5  NC_011333.1    1652982 38.89  Helicobacter pylori G27
    6  NC_011498.1    1673813 38.81  Helicobacter pylori P12
    7  NC_012973.1    1576758 39.16  Helicobacter pylori B38
  • nucmer -c 40 => ~200 alignments & 93-95% identity between genomes
  • SNPs are mostly substitutions
  • Alignment info (NC_000915 0cvg regions) :5-10% of genomes are unique
       .                      elem  min  q1   q2   q3   max    mean  n50   sum
    1  NC_000915-NC_000915    72    45   178  294  1890 10467  1013  1893  72976   #longest alignment has been removed
    2  NC_000915-NC_000921    197   2    81   203  495  17816  644   2146  126988
    3  NC_000915-NC_008086    151   3    103  242  894  26862  951   3103  143652
    4  NC_000915-NC_010698    206   2    115  283  706  12779  726   1941  149744
    5  NC_000915-NC_011333    138   2    111  260  695  7457   688   1941  95063
    6  NC_000915-NC_011498    157   2    83   185  565  5362   505   1357  79337
    7  NC_000915-NC_012973    140   2    108  239  526  37389  1018  5729  142568
  • NC_000915 vs NC_000915 : nucmer -c 40
 Align len
.                   elem       min    q1     q2     q3     max        mean       n50        sum
nucmer -c 20        484        20     21     26     82     10467      196        1892       95007
nucmer -c 40        72         45     178    294    1890   10467      1013       1893       72976
 Align %id
.                   elem       min    q1     q2     q3     max        mean       n50        sum
-c 20               484        63.72  91.89  100.00 100.00 100.00     95         100        46014.71
-c 40               72         76.71  85.06  92.86  99.92  100.00     92         93         6639.6
 Media:NC 000915-NC 000915.20.png, Media:NC 000915-NC 000915.40.png

NCBI SRA

Other

Assemblies

Wustl

   nl    assembly                                           ctgs  min  q1   q2    q3     max    mean  n50    sum      reads  0cvg_in_7_compl_genomes   
   1     HPKX_1039_AG0C1.scarf.assembled-29-21              233   100  318  1700  9639   77743  7085  19912  1650899  6.4m   674288   
   2     HPKX_1039_AG0C2.solexa.txt.assembled-23-12         420   101  341  1684  5301   36588  3914  9412   1644043  5.2M   729736   
   3     HPKX_1039_AG4C1.scarf.assembled-27-22              271   100  217  1421  8273   90368  6115  17093  1657230  6.0m   687296   
   4     HPKX_1039_AG4C2.solexa.txt.assembled-25-12         365   100  301  1595  5658   51523  4522  11890  1650547  6.8M   708607   

   5     HPKX_1172_AG0C1_090424.solexa.txt.assembled-25-30  217   107  557  3370  10683  58848  7099  15527  1540507  8.6M   1068848  
   6     HPKX_1172_AG0C2_2lanes.assembled-21-8              1170  100  264  717   1768   11444  1319  2661   1543511  7.2M   1110550  ?
   7     HPKX_1172_AG4C1_090424.solexa.txt.assembled-23-20  377   103  355  2178  6166   35180  4169  9160   1571948  8.5M   1106858  
   8     HPKX_1172_AG4C2.solexa.txt.assembled-25-15         317   100  274  1540  6256   37505  4987  14946  1581161  6.0M   812671   

   9     HPKX_1259_NL0C1.scarf.assembled-21-17              1704  100  264  598   1274   7953   936   1606   1595297  4.2m   963211   
   10    HPKX_1259_NL0C2.solexa.txt.assembled-23-12         410   102  240  928   4863   32792  3882  11295  1591864  3.6M   824474   
   11    HPKX_1259_NL4C1.scarf.assembled-27-23              283   100  224  1098  6814   98400  5634  18624  1594699  6.4m   797825   
   12    HPKX_1259_NL4C2.solexa.txt.assembled-23-12         455   102  222  874   4348   32792  3520  11010  1601950  6.3M   833155   

   13    HPKX_1379_NL0C1.scarf.assembled-27-22              295   100  230  1243  7551   59556  5539  15858  1634019  6.1m   730236   
   14    HPKX_1379_NL0C2.solexa.txt.assembled-23-12         416   100  216  1000  5177   53581  3931  11219  1635644  6.3M   754915   
   15    HPKX_1379_NL4C1.scarf.assembled-25-23              328   100  227  1084  6601   61090  4996  14203  1638925  5.0m   716540   
   16    HPKX_1379_NL4C2.solexa.txt.assembled-25-20         291   100  231  1501  6751   64227  5539  15080  1612046  4.6M   785276   

   17    HPKX_345_AG4C1.scarf.assembled-27-22               251   100  241  1272  8265   97643  6534  19718  1640151  4.5m   727208   
   18    HPKX_345_AG4C2                                     .     .    .    .     .      .      .     .      .        12.1M  .
   19    HPKX_345_NL0C1.scarf.assembled-25-30               305   100  243  1146  6718   59632  5360  15874  1634815  5.7m   759194   
   20    HPKX_345_NL0C2_090424.solexa.txt.assembled-25-26   283   100  254  2009  8300   59229  5629  13524  1593064  11.1M  1067288  

   21    HPKX_438_AG0C1.scarf.assembled-27-25               267   100  348  1710  8311   87876  6071  16918  1620975  5.8m   755933   
   22    HPKX_438_AG0C2.solexa.txt.assembled-23-18          407   102  396  1777  5455   31183  3963  8830   1613167  6.3M   804474   
   23    HPKX_438_CA4C1.scarf.assembled-27-26               237   100  348  1580  8856   97139  6845  19582  1622487  6.0m   742559   
   24    HPKX_438_CA4C2.solexa.txt.assembled-23-11          485   101  363  1502  4408   35471  3332  7779   1616183  4.1M   801123

CBCB

   nl  assembly                                             ctgs  min  q1   q2    q3     max    mean  n50    sum      reads
    2  HPKX_1039_AG0C2                                      335   45   91   1376  5910   51990  4822  14468  1615385
    4  HPKX_1039_AG4C2                                      371   45   76   297   5375   90459  4472  18459  1659148

    5  HPKX_1172_AG0C1                                      516   45   81   244   2838   44992  3075  12094  1587142
    6  HPKX_1172_AG0C2                                      1163  45   213  474   991    6591   740   1243   861451
    7  HPKX_1172_AG4C1                                      2214  45   148  287   530    3776   405   589    898322
    8  HPKX_1172_AG4C2                                      332   45   74   280   4800   50005  4741  20671  1574270

   10  HPKX_1259_NL0C2                                      414   45   72   295   3349   78486  3861  17825  1598491
   12  HPKX_1259_NL4C2                                      472   45   69   137   2017   59943  3381  18488  1596084

   14  HPKX_1379_NL0C2                                      358   45   75   191   3332   79512  4551  21347  1629546
   16  HPKX_1379_NL4C2                                      391   45   75   153   2314   79083  4184  22491  1635979

   24  HPKX_345_AG4C2                                       1098  45   244  724   1799   25718  1367  2745   1501014
   19  HPKX_345_NL0C2                                       1436  45   236  567   1260   8990   965   1796   1387059

   21  HPKX_438_AG0C2                                       410   45   90   562   4467   49129  3962  13859  1624766
   23  HPKX_438_CA4C2                                       398   45   161  1435  5137   37278  4068  12278  1619323

velvet_0.7.55:

  1. Fastq vs Fasta: no diffrence
  2. velvetg . -exp_cov auto



23 HPKX_438_CA4C2.solexa.txt.assembled-23-11

Reads

  • 4.1M Solexa 36bp unpaired
  • cvg =~ 80X
  • ~9% of the reads contain at least one N

Quality QC:

 .                  elem                                  min    q1     q2     q3     max        mean       n50        sum
 Ncount             4107397    3725799<=0    381598>0     0      0      0      0      35         1          34         5614614
 avgQuality         4107397    118980<=20    3988417>20   0      19     26     29     34         22         28         91690471
 Ncount==0 and avgQuality>=20 => 3013939 filtered reads (73%)
 pos                  elem       min    q1     q2     q3     max        mean       n50        sum
 0                    4107397    0      32     33     33     33         28         33         116108100
 1                    4107397    0      30     33     34     34         27         33         114845690
 5                    4107397    0      27     32     33     34         26         33         109832819
 10                   4107397    0      23     31     33     34         25         32         102860584
 20                   4107397    0      17     28     31     34         22         30         92139539
 30                   4107397    0      2      21     28     34         17         27         70231217
 32                   4107397    0      2      19     26     34         15         26         63156733
 35                   4107397    0      2      2      25     34         13         26         55261361

12mer counts: too much error???

 meryl -C -B -m 12 -s prefix.seq -o prefix.12mers
 meryl -Dh -s prefix.12mers | sort -nk2 -r | more
 1       1876075 0.3452  0.0196
 2       1009161 0.5308  0.0407
 ...
 9       36227   0.7772  0.1017
 10      25866   0.7819  0.1044
 48      20812   0.8729  0.2727  # read cvg ??? 
 49      20726   0.8768  0.2833
 ...

Velvet (all reads)

Ctg stats for different velveth hash_lengths:

 hash  #ctgs    min  q1   q2    q3    max    mean  n50    sum
 19    908      37   161  770   2289  21014  1732  3905   1572704
 21    457      41   84   580   4156  49037  3548  12777  1621652
 23    398      45   161  1435  5137  37278  4068  12278  1619323  (CBCB best*)
 27    769      53   341  1163  2731  18704  2109  4319   1622389

 ?     485      101  363  1502  4408  35471  3332  7779   1616183  (WUSTL)

CBCB best* read cvg =~ 23; repeats at higher cvg

 .     #ctgs    min  q1   q2    q3    max    mean  n50    sum
 cvg   398      13   21   23    25    139    30    25     .

Velvet (filtered reads)

Hash_len=23

Ctg stats

 filter           #ctgs    min  q1   q2    q3    max    mean  n50    sum          #reads     0cvg(all 6 genomes)
 all              398      45   161  1435  5137  37278  4068  12278  1619323      4107397    765345
 noN              420      45   115  1194  4898  37173  3863  11880  1622651      3725799    759343
 avgqual20+       424      45   150  1302  4797  36335  3828  11749  1623136      3069665    757697 
 noN.avgqual20+   453      45   137  1093  4394  36335  3586  11461  1624653      3013939    756829     !!! least seq missing

AMOScmp

Ref : NC_000915

Ctg stats:

 params                           #ctgs      min    q1     q2     q3     max        mean       n50        sum       #readsInCtgs
 -l 16 -c 32 -ovl 10              9533       36     59     96     168    3160       136        185        1302448   1,123,025 (~25%) 1,023,929:0SNP 154,958:1SNP 8,532:2SNP ...
 -l 8  -c 24 -ovl 10              4429       36     62     152    430    5518       350        806        1554095   2,438,762 (~50%) 1,159,669:0SNP 977,687:1SNP 422,738:2SNP ...
 -l 8  -c 24 -ovl 5               3880       36     61     158    492    5883       400        966        1553027   2,438,762 (~50%)

nucmer 0cvg stats:

params                            #gaps      min    q1     q2     q3     max        mean       n50        sum 
 -l 16 -c 32                      8708       2      10     19     41     9286       44         91         388608 
 -l 8  -c 24                      2650       2      8      17     45     2347       56         207        150099

24 HPKX_345_AG4C2

Reads

  • 12.1M Solexa 36bp unpaired
  • cvg =~ 120X ?

Velvet

Ctg stats :

 hash  #ctgs    min  q1   q2    q3    max    mean  n50    sum
 23    1098     45   244  724   1799  25718  1367  2745   1501014

NC_011498

NC_011498.1 1673813 38.81

Reads

  • 1.67M Simulated reads 36bp; unpaired; 100% correct;
  • the reads were generated by breaking the genome in 36bp segments (35bp ovl)=>36X cvg

Velvet

  • Ctg stats :
 hash                     #ctgs    min  q1   q2    q3    max    mean  n50    sum
 23                       292      45   67   164   3422  73108  5654  33268  1651121

Euler-sr

  • Ctg stats :
 vertex_size              #ctgs    min  q1   q2    q3    max    mean  n50    sum        #misassemblies  0cvg
 23                       366      24   36   92    931   83748  4596  41745  1682410    . 
 25                       343      26   39   98    1125  83752  5075  42087  1740988    4               27377
 27                       331      28   43  109    1125  83756  5016  41753  1660506    4               27392

AMOScmp

  • Ref : NC_000915
  • Ctg stats:
 params                                   #ctgs    min  q1   q2    q3    max    mean  n50    sum        #readsInCtgs    misassemblies(<95%length match)
 nucmer -l 16 -c 32 -ovl 10               8569     36   66   114   203   2897   164   231    1405371    836827  (~50%)  39 
 soap -v 5 -g 3 -s 12 -f 2; -ovl 10       1787     37   99   305   1129  14043  881   2283   1574554    1437078 (~85%)  69
 soap -v 5 -g 0 -s 12 -f 2; -ovl 10       1789     37   98   304   1128  14043  880   2283   1574516    1437077         64
 soap -v 3 -g 0 -s 12 -f 2; -ovl 10       3646     36   89   214   532   7184   424   857    1548982                    55
 soap -v 3 -g 0 -s 12 -f 2; -ovl 20       4957     36   81   174   389   4783   316   580    1567357    1353104         49
  • 0cvg stats
 params                                   #gaps    min  q1   q2    q3    max    mean  n50    sum
 nucmer -l 16 -c 32 -ovl 10               8026     2    8    15    34    5384   36    75     291959
 soap -v 5 -g 3 -s 12 -f 2; -ovl 10       1042     2    6    19    46    5355   93    746    97176

minimus* on velvet contigs

 .                                    ctgs       min    q1     q2     q3     max        mean       n50        sum            
 velvet                               292        45     67     164    3422   73108      5654       33268      1651121      
                                      ctgs+sing  min    q1     q2     q3     max        mean       n50        sum      misas.
 minimus2(delta-filter -1; OVL=20)    191        45     108    465    10421  117862     8631       33268      1648580  17(6)  # ctgs-vs-ctgs; OVL=20
 minimus3(delta-filter -q; OVL=20)    251        45     68     214    5486   73108      6573       33268      1650065  11()   # ctgs-vs-ref => ctgs-vs-ctgs
 minimus3(delta-filter -q; OVL=5)     191        45     71     227    9222   73108      8631       41743      1648698  6(1)
 minimus3(                OVL=20)     204        45     115    560    10394  117862     8072       33268      1646865  5(1)
 minimus3(                OVL=5)      172        45     134    611    12957  122309     9572       37367      1646538  8(4) 
 minimus3(all ref; OVL=20)            231        45     99     357    7424   117862     7138       33268      1648917  5(1)
 minimus3(all ref; OVL=5)             150        45     140    1177   15439  118850     10959      41743      1643998  15(7)