Unconfigured Ad

Collapse
X
 
  • Time
  • Show
Clear All
new posts
  • samhokin
    Member
    • Nov 2013
    • 20

    #1

    Tophat2 chose a match with a deletion over one without

    I'm a bit mystified by Tophat's (v2.1.0) choice of alignment in a case where I've got two similar genomes placed end to end (B73 and W22 maize) and I'm trying to map reads from a mixed (or non-mixed in this case) sample to their originating genome.

    So this case came up, where Tophat chose to assign a read to the B73 chromosome, even though it has an deletion in that case, while the same read maps to the corresponding W22 chromosome perfectly, without the deletion. Here's the BLAST comparison: "Chr2" is W22 chromosome 2, "2" is B73 chromosome 2. W22/Chr2 matches 100%, without gaps (e=1e-45); B73/2 has one gap (e=5e-44).

    Code:
    BLASTN 2.2.31+
    
    Reference: Zheng Zhang, Scott Schwartz, Lukas Wagner, and Webb
    Miller (2000), "A greedy algorithm for aligning DNA sequences", J
    Comput Biol 2000; 7(1-2):203-14.
    
    Database: B73v3.W22v3.genome.fa
               24 sequences; 4,458,629,333 total letters
    
    Query= DB775P1:321:C4J4FACXX:7:1304:6219:16563
    
    Length=101
                                                                          Score     E
    Sequences producing significant alignments:                          (Bits)  Value
    
      Chr2                                                                187     1e-45
      2 dna:chromosome chromosome:AGPv3:2:1:237917468:1                   182     5e-44
    
    > Chr2
    Length=251269698
    
     Score = 187 bits (101),  Expect = 1e-45
     Identities = 101/101 (100%), Gaps = 0/101 (0%)
     Strand=Plus/Plus
    
    Query  1          ACATAGTGCACCGATCACTGTTCTACTAGTATGGGCAGTAGAAGCAAAATGGTGGCGCGG  60
                      ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
    Sbjct  250221315  ACATAGTGCACCGATCACTGTTCTACTAGTATGGGCAGTAGAAGCAAAATGGTGGCGCGG  250221374
    
    Query  61         TCGCTTCTTTTCATGTTGACATCACATCCTGGAGTAGCCCT  101
                      |||||||||||||||||||||||||||||||||||||||||
    Sbjct  250221375  TCGCTTCTTTTCATGTTGACATCACATCCTGGAGTAGCCCT  250221415
    
    
    > 2 dna:chromosome chromosome:AGPv3:2:1:237917468:1
    Length=237917468
    
     Score = 182 bits (98),  Expect = 5e-44
     Identities = 101/102 (99%), Gaps = 1/102 (1%)
     Strand=Plus/Plus
    
    Query  1          ACATAGTGCACCGATCACTG-TTCTACTAGTATGGGCAGTAGAAGCAAAATGGTGGCGCG  59
                      |||||||||||||||||||| |||||||||||||||||||||||||||||||||||||||
    Sbjct  236204183  ACATAGTGCACCGATCACTGTTTCTACTAGTATGGGCAGTAGAAGCAAAATGGTGGCGCG  236204242
    
    Query  60         GTCGCTTCTTTTCATGTTGACATCACATCCTGGAGTAGCCCT  101
                      ||||||||||||||||||||||||||||||||||||||||||
    Sbjct  236204243  GTCGCTTCTTTTCATGTTGACATCACATCCTGGAGTAGCCCT  236204284
    
    Lambda      K        H
        1.33    0.621     1.12 
    
    Gapped
    Lambda      K        H
        1.28    0.460    0.850 
    
    Effective search space used: 325479892253
    
      Database: B73v3.W22v3.genome.fa
        Posted date:  Apr 30, 2016  9:09 AM
      Number of letters in database: 4,458,629,333
      Number of sequences in database:  24
    
    Matrix: blastn matrix 1 -2
    Gap Penalties: Existence: 0, Extension: 2.5
    The Tophat parameters are -g 1 -M -N 0 to enforce the choice of a single, perfect match. But Tophat should have chosen the W22 locus, not the B73 locus in this case.

    Any ideas why it chose the non-perfect match???
    Sam Hokin
    Computational Scientist, Carnegie and NCGR

Latest Articles

Collapse

  • SEQadmin2
    Multiomic Liquid Biopsy: What DNA/RNA Panels Reveal That DNA Alone Can’t
    by SEQadmin2



    Tumors contain complex mixtures of cell populations that evolve in response to treatment and host-specific factors. Tissue biopsies provide only static snapshots of a tumor’s state. But what if we could follow that evolution over time using a minimally invasive technique?

    Liquid biopsies can detect tumor-derived material circulating in the bloodstream, providing a way to study treatment response, detect emerging resistance, and discover new biomarkers. Circulating
    ...
    10-14-2026, 09:55 AM
  • SEQadmin2
    New Genomics Technologies Take Aim at Long-Standing Limits
    by SEQadmin2


    Researchers using sequencing and genomics tools often have to make trade-offs. They can choose between speed or scale, short reads or long-range information, or targeted panels or a view of the whole transcriptome. New technologies that have been released this year are built to address those tough choices.

    We asked six companies the same four questions to learn about their latest products. The new technologies bring a lot to the table, including rethinking sequencing
    ...
    09-28-2026, 10:25 AM

ad_right_rmr

Collapse

News

Collapse

Topics Statistics Last Post
Started by SEQadmin2, 10-22-2026, 11:13 AM
0 responses
37 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 10-22-2026, 10:48 AM
0 responses
28 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 10-15-2026, 10:52 AM
0 responses
41 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 10-14-2026, 05:39 AM
0 responses
45 views
0 reactions
Last Post SEQadmin2  
Working...