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Walk through tutorial

Joshua Gould edited this page Mar 23, 2021 · 16 revisions

Please be sure to have installed the CTAT-Mutations Pipeline software including setting up the CTAT genome library before running the pipeline.

Use the following command to run the pipeline, using the sample data provided in the testing/ subdirectory that's included with the ctat_mutations software.

   python /path/to/ctat_mutations \
       --left reads_1.fastq.gz \
       --right reads_2.fastq.gz \
       --sample_id test \
       --boosting_method none \
       --output varcalling.outdir 

Note, this step might take ~30 minutes to complete.

The demo data is a small set of reads and variants used for testing purposes. Let's take a look at cancer.tab output which lists all the cancer variants with respect to these small set of reads only:

CHROM  POS        REF  ALT  GENE     DP   QUAL               MQ  clinvar_sig                                    TUMOR                                                             TISSUE                              COSMIC_ID     FATHMM      chasmplus_pval  vest_pval  mupit_link
chr1   11130632   G    T    MTOR     59   745.6399999999999  60  NA                                             carcinoma_--_adenocarcinoma                                       lung                                COSV63879124  PATHOGENIC  0.0606          0.44247    "http://www.cravat.us/MuPIT_Interactive?gm=chr1:11130632"
chr1   11130740   A    C    MTOR     62   202.64             60  NA                                             carcinoma_--_adenocarcinoma                                       lung                                COSV63872139  PATHOGENIC  4.76e-05        0.04777    "http://www.cravat.us/MuPIT_Interactive?gm=chr1:11130740"
chr1   26774594   C    T    ARID1A   23   303.64             60  NA                                             malignant_melanoma_--_acral_lentiginous                           skin                                COSV61383982  PATHOGENIC  1.6e-06         0.01528    NA
chr1   114716123  C    T    NRAS     259  6233.06            60  "Pathogenic"                                   haematopoietic_neoplasm_--_acute_myeloid_leukaemia                haematopoietic_and_lymphoid_tissue  COSV54736416  PATHOGENIC  0               NA         "http://www.cravat.us/MuPIT_Interactive?gm=chr1:114716123"
chr2   177234087  T    A    NFE2L2   81   1812.06            60  NA                                             carcinoma_--_NS                                                   liver                               COSV67960411  PATHOGENIC  0               0.00344    "http://www.cravat.us/MuPIT_Interactive?gm=chr2:177234087"
chr2   201286575  T    G    CASP8    19   360.98             60  NA                                             carcinoma_--_adenocarcinoma                                       large_intestine                     COSV51852791  PATHOGENIC  3.2e-06         0.00536    "http://www.cravat.us/MuPIT_Interactive?gm=chr2:201286575"
chr2   208248388  C    T    IDH1     51   357.64             60  "Pathogenic"                                   glioma_--_oligodendroglioma_Grade_III                             central_nervous_system              COSV61615239  PATHOGENIC  0               0.02014    "http://www.cravat.us/MuPIT_Interactive?gm=chr2:208248388"
chr3   49375540   C    T    RHOA     646  5072.64            60  NA                                             carcinoma_--_adenocarcinoma                                       large_intestine                     COSV69041693  PATHOGENIC  3.15e-05        0.01042    "http://www.cravat.us/MuPIT_Interactive?gm=chr3:49375540"
chr3   179218303  G    A    PIK3CA   19   145.64             60  "Pathogenic/Likely-pathogenic"                 carcinoma_--_papillary_carcinoma                                  thyroid                             COSV55873239  PATHOGENIC  0               NA         "http://www.cravat.us/MuPIT_Interactive?gm=chr3:179218303"
chr5   68293360   C    T    PIK3R1   26   216.64             60  "Benign/Likely-benign"                         carcinoma_--_NS                                                   breast                              COSV57123112  PATHOGENIC  NA              NA         NA
chr5   112780896  G    A    APC      7    70.64              60  "Uncertain-significance"                       carcinoma_--_adenocarcinoma                                       stomach                             COSV57397660  PATHOGENIC  0.000105        0.05192    "http://www.cravat.us/MuPIT_Interactive?gm=chr5:112780896"
chr7   55191831   T    A    EGFR     190  675.6399999999999  60  "Pathogenic/Likely-pathogenic,-drug-response"  carcinoma_--_non_small_cell_carcinoma                             lung                                COSV51766344  PATHOGENIC  0               0.0082     "http://www.cravat.us/MuPIT_Interactive?gm=chr7:55191831"
chr12  25245350   C    A    KRAS     24   261.64             60  "Pathogenic"                                   carcinoma_--_adenocarcinoma                                       large_intestine                     COSV55497419  PATHOGENIC  0               0.01002    "http://www.cravat.us/MuPIT_Interactive?gm=chr12:25245350"
chr12  112450394  G    T    PTPN11   65   275.64             60  "Pathogenic"                                   glioma_--_astrocytoma_Grade_III                                   central_nervous_system              COSV61004869  PATHOGENIC  6.4e-07         0.01609    "http://www.cravat.us/MuPIT_Interactive?gm=chr12:112450394"
chr14  95103424   C    T    DICER1   10   252.06             60  "Benign/Likely-benign"                         carcinoma_--_endometrioid_carcinoma                               endometrium                         COSV58615839  PATHOGENIC  NA              NA         NA
chr16  3781283    C    T    CREBBP   23   79.64              60  NA                                             carcinoma_--_adenocarcinoma                                       large_intestine                     COSV52114432  PATHOGENIC  0.00543         0.92066    NA
chr16  68813455   C    A    CDH1     96   617.64             60  NA                                             carcinoma_--_squamous_cell_carcinoma                              oesophagus                          COSV55737831  PATHOGENIC  5.44e-05        0.00557    NA
chr17  7674221    G    A    TP53     26   737.06             60  "Pathogenic"                                   carcinoma_--_NS                                                   urinary_tract                       COSV52662035  PATHOGENIC  0               0.00071    "http://www.cravat.us/MuPIT_Interactive?gm=chr17:7674221"
chr17  39723967   T    C    ERBB2    80   967.64             60  "Likely-pathogenic"                            carcinoma_--_lobular_carcinoma                                    breast                              COSV54062780  PATHOGENIC  0               0.00293    "http://www.cravat.us/MuPIT_Interactive?gm=chr17:39723967"
chr19  12690014   T    G    FBXW9    42   156.64             60  NA                                             Ewings_sarcoma-peripheral_primitive_neuroectodermal_tumour_--_NS  bone                                COSV66709239  PATHOGENIC  0.306           0.15899    "http://www.cravat.us/MuPIT_Interactive?gm=chr19:12690014"
chr19  52216574   T    C    PPP2R1A  3    54.84              60  NA                                             NA                                                                NA                                  NA            NA          0.0178          0.03623    "http://www.cravat.us/MuPIT_Interactive?gm=chr19:52216574"
chr22  41170455   T    A    EP300    19   134.64             60  NA                                             glioma_--_astrocytoma_Grade_IV                                    central_nervous_system              COSV54331178  PATHOGENIC  0               NA         "http://www.cravat.us/MuPIT_Interactive?gm=chr22:41170455"

The corresponding interactive igv-js.html report is available as variants.HC_hard_cutoffs_applied.cancer.igvjs_viewer.html. Just right-click, save-as, and open up the html file in your web browser.