Tag Archives: RepeatMasker

Transposable Element Mapping – Crassostrea virginica Genome, Cvirginica_v300, using RepeatMasker 4.07

Per this GitHub issue, I’m IDing transposable elements (TEs) in the Crassostrea virginica genome.

Genome used:

I ran RepeatMasker (v4.07) with RepBase-20170127 and RMBlast 2.6.0 four times:

  1. Species = all

  2. Species = Crassostrea gigas (Pacific oyster)

  3. Species = Crassostrea virginica (Eastern oyster)

  4. Default settings (i.e. no species select – will use human genome).

The idea with running this with four different settings was to get a sense of how the analyses would differ with species specifications.

All runs were performed on roadrunner.

All commands were documented in a Jupyter Notebook (GitHub):

NOTE: RepeatMasker writes the desired output files (*.out, *.cat.gz, and *.gff) to the same directory that the genome is located in! If you conduct multiple runs with the same genome in the same directory, it will overwrite those files, as they are named using the genome assembly filename. Be sure to move files out of the genome directory after each run!


RESULTS:
RUN 1 (species – all)

Output folder:

Summary table (text):

Output table (GFF):

SUMMARY TABLE

==================================================
file name: Cvirginica_v300.fa       
sequences:            11
total length:  684741128 bp  (684675328 bp excl N/X-runs)
GC level:         34.83 %
bases masked:  113771462 bp ( 16.62 %)
==================================================
               number of      length   percentage
               elements*    occupied  of sequence
--------------------------------------------------
Retroelements        97003     27946871 bp    4.08 %
   SINEs:            48145      9242559 bp    1.35 %
   Penelope           1429       256929 bp    0.04 %
   LINEs:            27022     10570154 bp    1.54 %
    CRE/SLACS           28         2219 bp    0.00 %
     L2/CR1/Rex       2160       316660 bp    0.05 %
     R1/LOA/Jockey    3058       386611 bp    0.06 %
     R2/R4/NeSL        511       226938 bp    0.03 %
     RTE/Bov-B        7377      3276312 bp    0.48 %
     L1/CIN4          1331        95476 bp    0.01 %
   LTR elements:     21836      8134158 bp    1.19 %
     BEL/Pao          1807       936488 bp    0.14 %
     Ty1/Copia        3046       296183 bp    0.04 %
     Gypsy/DIRS1     12789      6060883 bp    0.89 %
       Retroviral     2369       152228 bp    0.02 %

DNA transposons     180693     29492426 bp    4.31 %
   hobo-Activator    12869      1114188 bp    0.16 %
   Tc1-IS630-Pogo    17233      2485049 bp    0.36 %
   En-Spm                0            0 bp    0.00 %
   MuDR-IS905            0            0 bp    0.00 %
   PiggyBac           2388       405926 bp    0.06 %
   Tourist/Harbinger  9302       992476 bp    0.14 %
   Other (Mirage,      238        15946 bp    0.00 %
    P-element, Transib)

Rolling-circles          0            0 bp    0.00 %

Unclassified:       137707     45460608 bp    6.64 %

Total interspersed repeats:   102899905 bp   15.03 %


Small RNA:           45243      9057873 bp    1.32 %

Satellites:           3852       760316 bp    0.11 %
Simple repeats:     203542      8946510 bp    1.31 %
Low complexity:      26205      1281043 bp    0.19 %
==================================================

* most repeats fragmented by insertions or deletions
  have been counted as one element
  Runs of >=20 X/Ns in query were excluded in % calcs


The query species was assumed to be root          
RepeatMasker Combined Database: Dfam_Consensus-20170127, RepBase-20170127
        
run with rmblastn version 2.6.0+


RUN 2 (species – Crassostrea gigas)

Output folder:

Summary table (text):

Output table (GFF):

SUMMARY TABLE

==================================================
file name: Cvirginica_v300.fa       
sequences:            11
total length:  684741128 bp  (684675328 bp excl N/X-runs)
GC level:         34.83 %
bases masked:   93923386 bp ( 13.72 %)
==================================================
               number of      length   percentage
               elements*    occupied  of sequence
--------------------------------------------------
Retroelements        26397     15008601 bp    2.19 %
   SINEs:                4          722 bp    0.00 %
   Penelope            675       190160 bp    0.03 %
   LINEs:            17645      8922188 bp    1.30 %
    CRE/SLACS            0            0 bp    0.00 %
     L2/CR1/Rex         70        39188 bp    0.01 %
     R1/LOA/Jockey       0            0 bp    0.00 %
     R2/R4/NeSL          4         5110 bp    0.00 %
     RTE/Bov-B        6194      2718955 bp    0.40 %
     L1/CIN4             0            0 bp    0.00 %
   LTR elements:      8748      6085691 bp    0.89 %
     BEL/Pao           933       788887 bp    0.12 %
     Ty1/Copia          47        82743 bp    0.01 %
     Gypsy/DIRS1      6819      4822734 bp    0.70 %
       Retroviral        0            0 bp    0.00 %

DNA transposons     163945     26422122 bp    3.86 %
   hobo-Activator     7742       720623 bp    0.11 %
   Tc1-IS630-Pogo    15615      2328538 bp    0.34 %
   En-Spm                0            0 bp    0.00 %
   MuDR-IS905            0            0 bp    0.00 %
   PiggyBac           2246       393498 bp    0.06 %
   Tourist/Harbinger  8431       876020 bp    0.13 %
   Other (Mirage,        0            0 bp    0.00 %
    P-element, Transib)

Rolling-circles          0            0 bp    0.00 %

Unclassified:       160681     41266796 bp    6.03 %

Total interspersed repeats:    82697519 bp   12.08 %


Small RNA:             214        40811 bp    0.01 %

Satellites:           1396       217317 bp    0.03 %
Simple repeats:     216869      9637447 bp    1.41 %
Low complexity:      27520      1418990 bp    0.21 %
==================================================

* most repeats fragmented by insertions or deletions
  have been counted as one element
  Runs of >=20 X/Ns in query were excluded in % calcs


The query species was assumed to be crassostrea gigas
RepeatMasker Combined Database: Dfam_Consensus-20170127, RepBase-20170127
        

RUN 3 (species – Crassostrea virginica)

Output folder:

Summary table (text):

Output table (GFF):

SUMMARY TABLE

==================================================
file name: Cvirginica_v300.fa       
sequences:            11
total length:  684741128 bp  (684675328 bp excl N/X-runs)
GC level:         34.83 %
bases masked:   46637065 bp ( 6.81 %)
==================================================
               number of      length   percentage
               elements*    occupied  of sequence
--------------------------------------------------
Retroelements        43139      8952068 bp    1.31 %
   SINEs:            43139      8952068 bp    1.31 %
   Penelope              0            0 bp    0.00 %
   LINEs:                0            0 bp    0.00 %
    CRE/SLACS            0            0 bp    0.00 %
     L2/CR1/Rex          0            0 bp    0.00 %
     R1/LOA/Jockey       0            0 bp    0.00 %
     R2/R4/NeSL          0            0 bp    0.00 %
     RTE/Bov-B           0            0 bp    0.00 %
     L1/CIN4             0            0 bp    0.00 %
   LTR elements:         0            0 bp    0.00 %
     BEL/Pao             0            0 bp    0.00 %
     Ty1/Copia           0            0 bp    0.00 %
     Gypsy/DIRS1         0            0 bp    0.00 %
       Retroviral        0            0 bp    0.00 %

DNA transposons       3538      1564942 bp    0.23 %
   hobo-Activator        0            0 bp    0.00 %
   Tc1-IS630-Pogo        0            0 bp    0.00 %
   En-Spm                0            0 bp    0.00 %
   MuDR-IS905            0            0 bp    0.00 %
   PiggyBac              0            0 bp    0.00 %
   Tourist/Harbinger     0            0 bp    0.00 %
   Other (Mirage,        0            0 bp    0.00 %
    P-element, Transib)

Rolling-circles          0            0 bp    0.00 %

Unclassified:        65151     23982146 bp    3.50 %

Total interspersed repeats:    34499156 bp    5.04 %


Small RNA:           43353      8992879 bp    1.31 %

Satellites:              1          222 bp    0.00 %
Simple repeats:     232627     10544162 bp    1.54 %
Low complexity:      29762      1561018 bp    0.23 %
==================================================

* most repeats fragmented by insertions or deletions
  have been counted as one element
  Runs of >=20 X/Ns in query were excluded in % calcs


The query species was assumed to be crassostrea virginica
RepeatMasker Combined Database: Dfam_Consensus-20170127, RepBase-20170127
        
run with rmblastn version 2.6.0+

RUN 4 (default settings – human genome)

Output folder:

Summary table (text):

Output table (GFF):

SUMMARY TABLE

==================================================
file name: Cvirginica_v300.fa       
sequences:            11
total length:  684741128 bp  (684675328 bp excl N/X-runs)
GC level:         34.83 %
bases masked:   13461422 bp ( 1.97 %)
==================================================
               number of      length   percentage
               elements*    occupied  of sequence
--------------------------------------------------
SINEs:             2056       120820 bp    0.02 %
      ALUs            0            0 bp    0.00 %
      MIRs          240        14635 bp    0.00 %

LINEs:             3408       331585 bp    0.05 %
      LINE1         240        16835 bp    0.00 %
      LINE2         728        69177 bp    0.01 %
      L3/CR1       1369       135234 bp    0.02 %

LTR elements:       704       236625 bp    0.03 %
      ERVL           14          944 bp    0.00 %
      ERVL-MaLRs     12          892 bp    0.00 %
      ERV_classI    272        36695 bp    0.01 %
      ERV_classII     4          206 bp    0.00 %

DNA elements:      1088       100026 bp    0.01 %
     hAT-Charlie     27         1543 bp    0.00 %
     TcMar-Tigger   142         9891 bp    0.00 %

Unclassified:        57         6096 bp    0.00 %

Total interspersed repeats:   795152 bp    0.12 %


Small RNA:         3698       279669 bp    0.04 %

Satellites:          73         5524 bp    0.00 %
Simple repeats:  247957     10848509 bp    1.58 %
Low complexity:   30084      1536314 bp    0.22 %
==================================================

* most repeats fragmented by insertions or deletions
  have been counted as one element
  Runs of >=20 X/Ns in query were excluded in % calcs


The query species was assumed to be homo sapiens  
RepeatMasker Combined Database: Dfam_Consensus-20170127, RepBase-20170127
        
run with rmblastn version 2.6.0+

Transposable Element Mapping – Olympia Oyster Genome Assembly, Olurida_v081, using RepeatMasker 4.07

I previously performed this analysis using a different version of our Ostrea lurida genome assembly. Steven asked that I repeat the analysis with a modified version of the genome assembly (Olurida_v081) – only has contigs >1000bp in length.

Genome used: Olurida_v081

I ran RepeatMasker (v4.07) with RepBase-20170127 and RMBlast 2.6.0 four times:

  1. Default settings (i.e. no species select – will use human genome).

  2. Species = Crassostrea gigas (Pacific oyster)

  3. Species = Crassostrea virginica (Eastern oyster)

  4. Species = Ostrea lurida (Olympia oyster)

The idea was to get a sense of how the analyses would differ with species specifications. However, it’s likely that the only species setting that will make any difference will be Run #2 (Crassostrea gigas).

The reason I say this is that RepeatMasker has a built in tool to query which species are available in the RepBase database (e.g.):

RepeatMasker-4.0.7/util/queryRepeatDatabase.pl -species "crassostrea virginica" -stat

Here’s a very brief overview of what that yields:

  • Crassotrea gigas: 792 specific repeats

  • Crassostrea virginica: 4 Crassostrea virginica specific repeats

  • Ostrea lurida: 0 Ostrea lurida specific repeats

All runs were performed on roadrunner.

All commands were documented in a Jupyter Notebook (GitHub):

NOTE: RepeatMasker writes the desired output files (*.out, *.cat.gz, and *.gff) to the same directory that the genome is located in! If you conduct multiple runs with the same genome in the same directory, it will overwrite those files, as they are named using the genome assembly filename.


RESULTS:
RUN 1 (default settings – human genome)

Output folder:

Summary table (text):

Output table (GFF):

SUMMARY TABLE

==================================================
file name: Olurida_v081.fa          
sequences:        159429
total length: 1140787867 bp  (1077373535 bp excl N/X-runs)
GC level:         36.58 %
bases masked:   17954347 bp ( 1.67 %)
==================================================
               number of      length   percentage
               elements*    occupied  of sequence
--------------------------------------------------
SINEs:            16599       978030 bp    0.09 %
      ALUs            1          292 bp    0.00 %
      MIRs          937        72873 bp    0.01 %

LINEs:             3279       752631 bp    0.07 %
      LINE1         172        10882 bp    0.00 %
      LINE2         646        67827 bp    0.01 %
      L3/CR1        659        60327 bp    0.01 %

LTR elements:       569       127808 bp    0.01 %
      ERVL           32         1949 bp    0.00 %
      ERVL-MaLRs     10          490 bp    0.00 %
      ERV_classI    165        17699 bp    0.00 %
      ERV_classII    26         1590 bp    0.00 %

DNA elements:      1911       161957 bp    0.02 %
     hAT-Charlie     74         4216 bp    0.00 %
     TcMar-Tigger   584        24985 bp    0.00 %

Unclassified:        78         9834 bp    0.00 %

Total interspersed repeats:  2030260 bp    0.19 %


Small RNA:         5592       409456 bp    0.04 %

Satellites:         117        21278 bp    0.00 %
Simple repeats:  270784     12935570 bp    1.20 %
Low complexity:   42130      2568284 bp    0.24 %
==================================================

* most repeats fragmented by insertions or deletions
  have been counted as one element
  Runs of >=20 X/Ns in query were excluded in % calcs


The query species was assumed to be homo sapiens  
RepeatMasker Combined Database: Dfam_Consensus-20170127, RepBase-20170127
        
run with rmblastn version 2.6.0+

RUN 2 (species – Crassostrea gigas)

Output folder:

Summary table (text):

Output table (GFF):

SUMMARY TABLE

file name: Olurida_v081.fa          
sequences:        159429
total length: 1140787867 bp  (1077373535 bp excl N/X-runs)
GC level:         36.58 %
bases masked:  152816516 bp ( 14.18 %)
==================================================
               number of      length   percentage
               elements*    occupied  of sequence
--------------------------------------------------
Retroelements       193250     67253771 bp    6.24 %
   SINEs:             2087       284274 bp    0.03 %
   Penelope         158576     56080082 bp    5.21 %
   LINEs:           179430     61300904 bp    5.69 %
    CRE/SLACS            0            0 bp    0.00 %
     L2/CR1/Rex        675       348273 bp    0.03 %
     R1/LOA/Jockey       0            0 bp    0.00 %
     R2/R4/NeSL          7        10781 bp    0.00 %
     RTE/Bov-B        7051      1827344 bp    0.17 %
     L1/CIN4             0            0 bp    0.00 %
   LTR elements:     11733      5668593 bp    0.53 %
     BEL/Pao          1517       871288 bp    0.08 %
     Ty1/Copia          78        72481 bp    0.01 %
     Gypsy/DIRS1      9151      4445789 bp    0.41 %
       Retroviral        0            0 bp    0.00 %

DNA transposons     233691     33727339 bp    3.13 %
   hobo-Activator    17578      1886743 bp    0.18 %
   Tc1-IS630-Pogo    39184      6403235 bp    0.59 %
   En-Spm                0            0 bp    0.00 %
   MuDR-IS905            0            0 bp    0.00 %
   PiggyBac           7261      1003937 bp    0.09 %
   Tourist/Harbinger  8635       823434 bp    0.08 %
   Other (Mirage,        0            0 bp    0.00 %
    P-element, Transib)

Rolling-circles          0            0 bp    0.00 %

Unclassified:       157855     36675484 bp    3.40 %

Total interspersed repeats:   137656594 bp   12.78 %


Small RNA:             222        72690 bp    0.01 %

Satellites:           6260      1238331 bp    0.11 %
Simple repeats:     241081     11662466 bp    1.08 %
Low complexity:      38915      2347827 bp    0.22 %
==================================================

* most repeats fragmented by insertions or deletions
  have been counted as one element
  Runs of >=20 X/Ns in query were excluded in % calcs


The query species was assumed to be crassostrea gigas
RepeatMasker Combined Database: Dfam_Consensus-20170127, RepBase-20170127
        
run with rmblastn version 2.6.0+

RUN 3 (species – Crassostrea virginica)

Output folder:

Summary table (text):

Output table (GFF):

SUMMARY TABLE

file name: Olurida_v081.fa          
sequences:        159429
total length: 1140787867 bp  (1077373535 bp excl N/X-runs)
GC level:         36.58 %
bases masked:   36996910 bp ( 3.43 %)
==================================================
               number of      length   percentage
               elements*    occupied  of sequence
--------------------------------------------------
Retroelements        59806      9886111 bp    0.92 %
   SINEs:            59806      9886111 bp    0.92 %
   Penelope              0            0 bp    0.00 %
   LINEs:                0            0 bp    0.00 %
    CRE/SLACS            0            0 bp    0.00 %
     L2/CR1/Rex          0            0 bp    0.00 %
     R1/LOA/Jockey       0            0 bp    0.00 %
     R2/R4/NeSL          0            0 bp    0.00 %
     RTE/Bov-B           0            0 bp    0.00 %
     L1/CIN4             0            0 bp    0.00 %
   LTR elements:         0            0 bp    0.00 %
     BEL/Pao             0            0 bp    0.00 %
     Ty1/Copia           0            0 bp    0.00 %
     Gypsy/DIRS1         0            0 bp    0.00 %
       Retroviral        0            0 bp    0.00 %

DNA transposons       8720      2230426 bp    0.21 %
   hobo-Activator        0            0 bp    0.00 %
   Tc1-IS630-Pogo        0            0 bp    0.00 %
   En-Spm                0            0 bp    0.00 %
   MuDR-IS905            0            0 bp    0.00 %
   PiggyBac              0            0 bp    0.00 %
   Tourist/Harbinger     0            0 bp    0.00 %
   Other (Mirage,        0            0 bp    0.00 %
    P-element, Transib)

Rolling-circles          0            0 bp    0.00 %

Unclassified:        47005      9434652 bp    0.88 %

Total interspersed repeats:    21551189 bp    2.00 %


Small RNA:           60030      9959172 bp    0.92 %

Satellites:              8         5100 bp    0.00 %
Simple repeats:     259134     12795379 bp    1.19 %
Low complexity:      42184      2581162 bp    0.24 %
==================================================

* most repeats fragmented by insertions or deletions
  have been counted as one element
  Runs of >=20 X/Ns in query were excluded in % calcs


The query species was assumed to be crassostrea virginica
RepeatMasker Combined Database: Dfam_Consensus-20170127, RepBase-20170127
        
run with rmblastn version 2.6.0+

RUN 4 (species – Ostrea lurida)

Output folder:

Summary table (text):

Output table (GFF):

SUMMARY TABLE

==================================================
file name: Olurida_v081.fa          
sequences:        159429
total length: 1140787867 bp  (1077373535 bp excl N/X-runs)
GC level:         36.58 %
bases masked:   15918797 bp ( 1.48 %)
==================================================
               number of      length   percentage
               elements*    occupied  of sequence
--------------------------------------------------
Retroelements            0            0 bp    0.00 %
   SINEs:                0            0 bp    0.00 %
   Penelope              0            0 bp    0.00 %
   LINEs:                0            0 bp    0.00 %
    CRE/SLACS            0            0 bp    0.00 %
     L2/CR1/Rex          0            0 bp    0.00 %
     R1/LOA/Jockey       0            0 bp    0.00 %
     R2/R4/NeSL          0            0 bp    0.00 %
     RTE/Bov-B           0            0 bp    0.00 %
     L1/CIN4             0            0 bp    0.00 %
   LTR elements:         0            0 bp    0.00 %
     BEL/Pao             0            0 bp    0.00 %
     Ty1/Copia           0            0 bp    0.00 %
     Gypsy/DIRS1         0            0 bp    0.00 %
       Retroviral        0            0 bp    0.00 %

DNA transposons          0            0 bp    0.00 %
   hobo-Activator        0            0 bp    0.00 %
   Tc1-IS630-Pogo        0            0 bp    0.00 %
   En-Spm                0            0 bp    0.00 %
   MuDR-IS905            0            0 bp    0.00 %
   PiggyBac              0            0 bp    0.00 %
   Tourist/Harbinger     0            0 bp    0.00 %
   Other (Mirage,        0            0 bp    0.00 %
    P-element, Transib)

Rolling-circles          0            0 bp    0.00 %

Unclassified:            3          189 bp    0.00 %

Total interspersed repeats:         189 bp    0.00 %


Small RNA:             224        73061 bp    0.01 %

Satellites:              8         5100 bp    0.00 %
Simple repeats:     273098     13256460 bp    1.23 %
Low complexity:      42443      2592212 bp    0.24 %
==================================================

* most repeats fragmented by insertions or deletions
  have been counted as one element
  Runs of >=20 X/Ns in query were excluded in % calcs


The query species was assumed to be ostrea lurida 
RepeatMasker Combined Database: Dfam_Consensus-20170127, RepBase-20170127
        
run with rmblastn version 2.6.0+



Transposable Element Mapping – Crassostrea virginica NCBI Genome Assembly using RepeatMasker 4.07

Genome used: NCBI GCA_002022765.4_C_virginica-3.0

I ran RepeatMasker (v4.07) with RepBase-20170127 and RMBlast 2.6.0 with species set to Crassotrea virginica.

All commands were documented in a Jupyter Notebook (GitHub):


RESULTS:

Output folder:

Output table (GFF):

Summary table (text):

==================================================
file name: GCF_002022765.2_C_virginica-3.0_genomic.fasta
sequences:            11
total length:  684741128 bp  (684675328 bp excl N/X-runs)
GC level:         34.83 %
bases masked:   46637065 bp ( 6.81 %)
==================================================
               number of      length   percentage
               elements*    occupied  of sequence
--------------------------------------------------
Retroelements        43139      8952068 bp    1.31 %
   SINEs:            43139      8952068 bp    1.31 %
   Penelope              0            0 bp    0.00 %
   LINEs:                0            0 bp    0.00 %
    CRE/SLACS            0            0 bp    0.00 %
     L2/CR1/Rex          0            0 bp    0.00 %
     R1/LOA/Jockey       0            0 bp    0.00 %
     R2/R4/NeSL          0            0 bp    0.00 %
     RTE/Bov-B           0            0 bp    0.00 %
     L1/CIN4             0            0 bp    0.00 %
   LTR elements:         0            0 bp    0.00 %
     BEL/Pao             0            0 bp    0.00 %
     Ty1/Copia           0            0 bp    0.00 %
     Gypsy/DIRS1         0            0 bp    0.00 %
       Retroviral        0            0 bp    0.00 %

DNA transposons       3538      1564942 bp    0.23 %
   hobo-Activator        0            0 bp    0.00 %
   Tc1-IS630-Pogo        0            0 bp    0.00 %
   En-Spm                0            0 bp    0.00 %
   MuDR-IS905            0            0 bp    0.00 %
   PiggyBac              0            0 bp    0.00 %
   Tourist/Harbinger     0            0 bp    0.00 %
   Other (Mirage,        0            0 bp    0.00 %
    P-element, Transib)

Rolling-circles          0            0 bp    0.00 %

Unclassified:        65151     23982146 bp    3.50 %

Total interspersed repeats:    34499156 bp    5.04 %


Small RNA:           43353      8992879 bp    1.31 %

Satellites:              1          222 bp    0.00 %
Simple repeats:     232627     10544162 bp    1.54 %
Low complexity:      29762      1561018 bp    0.23 %
==================================================

* most repeats fragmented by insertions or deletions
  have been counted as one element
  Runs of >=20 X/Ns in query were excluded in % calcs


The query species was assumed to be crassostrea virginica
RepeatMasker Combined Database: Dfam_Consensus-20170127, RepBase-20170127
        
run with rmblastn version 2.6.0+

Transposable Element Mapping – Olympia Oyster Genome Assembly using RepeatMasker 4.07

Steven wanted transposable elements (TEs) in the Olympia oyster genome identified.

After some minor struggles, I was able to get RepeatMasker installed on on both of our Apple Xserves (emu & roadrunner; running Ubuntu 16.04LTS).

Genome used: pbjelly_sjw_01

I ran RepeatMasker (v4.07) with RepBase-20170127 and RMBlast 2.6.0 four times:

  1. Default settings (i.e. no species select – will use human genome).

  2. Species = Crassostrea gigas (Pacific oyster)

  3. Species = Crassostrea virginica (Eastern oyster)

  4. Species = Ostrea lurida (Olympia oyster)

The idea was to get a sense of how the analyses would differ with species specifications. However, it’s likely that the only species setting that will make any difference will be Run #2 (Crassostrea gigas).

The reason I say this is that RepeatMasker has a built in tool to query which species are available in the RepBase database (e.g.):

RepeatMasker-4.0.7/util/queryRepeatDatabase.pl -species "crassostrea virginica" -stat

Here’s a very brief overview of what that yields:

  • Crassotrea gigas: 792 specific repeats

  • Crassostrea virginica: 4 Crassostrea virginica specific repeats

  • Ostrea lurida: 0 Ostrea lurida specific repeats

All runs were performed on roadrunner.

All commands were documented in a Jupyter Notebook (GitHub):

NOTE: RepeatMasker writes the desired output files (*.out, *.cat.gz, and *.gff) to the same directory that the genome is located in! If you conduct multiple runs with the same genome in the same directory, it will overwrite those files, as they are named using the genome assembly filename.


RESULTS:
RUN 1 (default settings – human genome)

Output folder:

Summary table (text):

Output table (GFF):

SUMMARY TABLE

==================================================
file name: jelly.out.fasta          
sequences:        696946
total length: 1253001795 bp  (1172226648 bp excl N/X-runs)
GC level:         36.51 %
bases masked:   20002806 bp ( 1.71 %)
==================================================
               number of      length   percentage
               elements*    occupied  of sequence
--------------------------------------------------
SINEs:            17794      1061170 bp    0.09 %
      ALUs          363        31340 bp    0.00 %
      MIRs         1166        92129 bp    0.01 %

LINEs:             4456       888114 bp    0.08 %
      LINE1         976       103929 bp    0.01 %
      LINE2         813        82891 bp    0.01 %
      L3/CR1        699        63627 bp    0.01 %

LTR elements:      1187       199118 bp    0.02 %
      ERVL          155        15828 bp    0.00 %
      ERVL-MaLRs    200        20737 bp    0.00 %
      ERV_classI    379        42833 bp    0.00 %
      ERV_classII    66         6896 bp    0.00 %

DNA elements:      2290       196866 bp    0.02 %
     hAT-Charlie    190        15468 bp    0.00 %
     TcMar-Tigger   732        37473 bp    0.00 %

Unclassified:       101        12946 bp    0.00 %

Total interspersed repeats:  2358214 bp    0.20 %


Small RNA:         5954       433422 bp    0.04 %

Satellites:         366        55705 bp    0.00 %
Simple repeats:  310641     14322152 bp    1.22 %
Low complexity:   47381      2844279 bp    0.24 %
==================================================

* most repeats fragmented by insertions or deletions
  have been counted as one element
  Runs of >=20 X/Ns in query were excluded in % calcs


The query species was assumed to be homo sapiens  
RepeatMasker Combined Database: Dfam_Consensus-20170127, RepBase-20170127
        
run with rmblastn version 2.6.0+

RUN 2 (species – Crassostrea gigas)

Output folder:

Summary table (text):

Output table (GFF):

SUMMARY TABLE

==================================================
file name: jelly.out.fasta          
sequences:        696946
total length: 1253001795 bp  (1172226648 bp excl N/X-runs)
GC level:         36.51 %
bases masked:  160759267 bp ( 13.71 %)
==================================================
               number of      length   percentage
               elements*    occupied  of sequence
--------------------------------------------------
Retroelements       213132     69887654 bp    5.96 %
   SINEs:             2374       311974 bp    0.03 %
   Penelope         171792     57862186 bp    4.94 %
   LINEs:           195605     63430615 bp    5.41 %
    CRE/SLACS            0            0 bp    0.00 %
     L2/CR1/Rex        731       357995 bp    0.03 %
     R1/LOA/Jockey       0            0 bp    0.00 %
     R2/R4/NeSL         13        11377 bp    0.00 %
     RTE/Bov-B        8085      1948581 bp    0.17 %
     L1/CIN4             0            0 bp    0.00 %
   LTR elements:     15153      6145065 bp    0.52 %
     BEL/Pao          2119       955773 bp    0.08 %
     Ty1/Copia         101        75372 bp    0.01 %
     Gypsy/DIRS1     11776      4815361 bp    0.41 %
       Retroviral        0            0 bp    0.00 %

DNA transposons     256292     35689117 bp    3.04 %
   hobo-Activator    19847      2059651 bp    0.18 %
   Tc1-IS630-Pogo    43269      6806311 bp    0.58 %
   En-Spm                0            0 bp    0.00 %
   MuDR-IS905            0            0 bp    0.00 %
   PiggyBac           7935      1060296 bp    0.09 %
   Tourist/Harbinger  9503       887332 bp    0.08 %
   Other (Mirage,        0            0 bp    0.00 %
    P-element, Transib)

Rolling-circles          0            0 bp    0.00 %

Unclassified:       174943     38299211 bp    3.27 %

Total interspersed repeats:   143875982 bp   12.27 %


Small RNA:             280        78768 bp    0.01 %

Satellites:           7383      1362194 bp    0.12 %
Simple repeats:     278809     12982714 bp    1.11 %
Low complexity:      44078      2622506 bp    0.22 %
==================================================

* most repeats fragmented by insertions or deletions
  have been counted as one element
  Runs of >=20 X/Ns in query were excluded in % calcs


The query species was assumed to be crassostrea gigas
RepeatMasker Combined Database: Dfam_Consensus-20170127, RepBase-20170127
        
run with rmblastn version 2.6.0+

RUN 3 (species – Crassostrea virginica)

Output folder:

Summary table (text):

Output table (GFF):

SUMMARY TABLE

==================================================
file name: jelly.out.fasta          
sequences:        696946
total length: 1253001795 bp  (1172226648 bp excl N/X-runs)
GC level:         36.51 %
bases masked:   39598953 bp ( 3.38 %)
==================================================
               number of      length   percentage
               elements*    occupied  of sequence
--------------------------------------------------
Retroelements        63882     10327611 bp    0.88 %
   SINEs:            63882     10327611 bp    0.88 %
   Penelope              0            0 bp    0.00 %
   LINEs:                0            0 bp    0.00 %
    CRE/SLACS            0            0 bp    0.00 %
     L2/CR1/Rex          0            0 bp    0.00 %
     R1/LOA/Jockey       0            0 bp    0.00 %
     R2/R4/NeSL          0            0 bp    0.00 %
     RTE/Bov-B           0            0 bp    0.00 %
     L1/CIN4             0            0 bp    0.00 %
   LTR elements:         0            0 bp    0.00 %
     BEL/Pao             0            0 bp    0.00 %
     Ty1/Copia           0            0 bp    0.00 %
     Gypsy/DIRS1         0            0 bp    0.00 %
       Retroviral        0            0 bp    0.00 %

DNA transposons       9433      2307292 bp    0.20 %
   hobo-Activator        0            0 bp    0.00 %
   Tc1-IS630-Pogo        0            0 bp    0.00 %
   En-Spm                0            0 bp    0.00 %
   MuDR-IS905            0            0 bp    0.00 %
   PiggyBac              0            0 bp    0.00 %
   Tourist/Harbinger     0            0 bp    0.00 %
   Other (Mirage,        0            0 bp    0.00 %
    P-element, Transib)

Rolling-circles          0            0 bp    0.00 %

Unclassified:        51558      9836468 bp    0.84 %

Total interspersed repeats:    22471371 bp    1.92 %


Small RNA:           64164     10406776 bp    0.89 %

Satellites:             10         5985 bp    0.00 %
Simple repeats:     298612     14185090 bp    1.21 %
Low complexity:      47510      2866522 bp    0.24 %
==================================================

* most repeats fragmented by insertions or deletions
  have been counted as one element
  Runs of >=20 X/Ns in query were excluded in % calcs


The query species was assumed to be crassostrea virginica
RepeatMasker Combined Database: Dfam_Consensus-20170127, RepBase-20170127
        
run with rmblastn version 2.6.0+

RUN 4 (species – Ostrea lurida)

Output folder:

Summary table (text):

Output table (GFF):

SUMMARY TABLE

==================================================
file name: jelly.out.fasta          
sequences:        696946
total length: 1253001795 bp  (1172226648 bp excl N/X-runs)
GC level:         36.51 %
bases masked:   17617763 bp ( 1.50 %)
==================================================
               number of      length   percentage
               elements*    occupied  of sequence
--------------------------------------------------
Retroelements            0            0 bp    0.00 %
   SINEs:                0            0 bp    0.00 %
   Penelope              0            0 bp    0.00 %
   LINEs:                0            0 bp    0.00 %
    CRE/SLACS            0            0 bp    0.00 %
     L2/CR1/Rex          0            0 bp    0.00 %
     R1/LOA/Jockey       0            0 bp    0.00 %
     R2/R4/NeSL          0            0 bp    0.00 %
     RTE/Bov-B           0            0 bp    0.00 %
     L1/CIN4             0            0 bp    0.00 %
   LTR elements:         0            0 bp    0.00 %
     BEL/Pao             0            0 bp    0.00 %
     Ty1/Copia           0            0 bp    0.00 %
     Gypsy/DIRS1         0            0 bp    0.00 %
       Retroviral        0            0 bp    0.00 %

DNA transposons          0            0 bp    0.00 %
   hobo-Activator        0            0 bp    0.00 %
   Tc1-IS630-Pogo        0            0 bp    0.00 %
   En-Spm                0            0 bp    0.00 %
   MuDR-IS905            0            0 bp    0.00 %
   PiggyBac              0            0 bp    0.00 %
   Tourist/Harbinger     0            0 bp    0.00 %
   Other (Mirage,        0            0 bp    0.00 %
    P-element, Transib)

Rolling-circles          0            0 bp    0.00 %

Unclassified:            3          189 bp    0.00 %

Total interspersed repeats:         189 bp    0.00 %


Small RNA:             282        79165 bp    0.01 %

Satellites:             10         5985 bp    0.00 %
Simple repeats:     313082     14662647 bp    1.25 %
Low complexity:      47785      2878201 bp    0.25 %
==================================================

* most repeats fragmented by insertions or deletions
  have been counted as one element
  Runs of >=20 X/Ns in query were excluded in % calcs


The query species was assumed to be ostrea lurida 
RepeatMasker Combined Database: Dfam_Consensus-20170127, RepBase-20170127
        
run with rmblastn version 2.6.0+

Software Installation – RepeatMasker v4.0.7 on Emu/Roadrunner Continued

After yesterday’s difficulties getting RMblast to compile, I deleted the folder and went through the build process again.

This time it worked, but it did not put rmblastn in the specified location (/home/shared/rmblast).

This fact took me a fair amount of time to figure out. Finally, after a couple of different re-builds, I ran find to see if rmblastn existed somewhere I wasn’t looking:

Additionally, I couldn’t find the location of the various BLAST executables. Some internet sleuthing led me to the NCBI page on installing BLAST+ from source, which indicates that the executables are stored in:

ncbi-blast-VERSION+-src/c++/ReleaseMT/bin/

How intuitive! /s

In order to improve readability and usability of the /home/shared/ directory, I renamed the /home/shared/rmblast directory to reflect the BLAST version and created a symbolic link in that directory to the rmlbastn executable:

Symbolic link to RMBLAST

Initiate RepeatMasker configuration


Confirm perl install location:


Confirm RepeatMasker install location:


Specify TRF install location:


Hmmm, TRF error. Looking for file called trf:


Renamed TRF file to trf and now it’s automatically found:


Set RMBlast as search engine:


Set RMBlast install location:


Set RMBlast as default search engine:


Confirmation of RMBlast as default search engine and successful installation of RepeatMasker:


Software Installation – RepeatMasker v4.0.7 on Emu/Roadrunner

Steven asked that I re-run some Olympia oyster transposable elements analysis using RepeatMasker and a newer version of our Olympia oyster genome assembly.

Installed the software on both of the Apple Xserves (Emu and Roadrunner) running Ubuntu 16.04.

Followed the instructions outlined here:

Starting with the prerequisites:

1. Download and install RMBlast

  • NCBI Blast 2.6.0 source

  • isb 2.6.0 patch

Unfortunately, the make command continually failed:

cd /home/shared/ncbi-blast-2.6.0+-src/c++
make

While trying to troubleshoot this issue, continued with the other prerequisites:

2. Downloaded Tandem Repeat Finder v.4.09

  • Saved file (trf409.linux64) to /home/shared/bin. NOTE: /home/shared/bin is part of the system PATH. See the /etc/environment file.

  • Changed permissions to be executable:

sudo chmod 775 trf409.linux64

3. Downloaded RepBase RepeatMasker Edition 20170127 (NOTE: This requires registration in order to obtain a username/password to download the file).

Installed RepeatMasker:

4. Downloaded RepeatMasker 4.0.7

  • Saved to /home/shared/RepeatMasker-4.0.7

5. Installed RepBase RepeatMasker Edition 20170127 in /home/shared//home/shared/RepeatMasker-4.0.7/Libraries

Currently re-building RMBlast and it takes forever… Will report back when I have it running.