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LeftAlignIndels Alignments added out of order... Offending records are at [chr7:55268881] and [chr7:55268881]



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    Hi James Covino

    I am not sure if you could solve your issue but let me ask. Have you observed any weird reads within bedtools intersect output via IGV or just samtools view command ?

    Would you be able to share any images of the region? 

    Also I noticed that you are trying to run these tools on a Apple Silicon Mac therefore you are receiving many error messages with response to library incompatibilities. Can you try running these in a compatible x86_64 compute environment and see if the issue still persists?


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    James Covino

    Hi @SkyWarrior,  
    Thanks for the reply.  
    I ran this on my personal Mac (Silicon) but we saw and duplicated this error on our deployed instance (Intel(R) Xeon(R) Platinum 8375C CPU @ 2.90GHz).   
    I'm attaching a sam with two alignments that result in the error from LeftAlignIndels. It appears that the first alignment is simplified into 56S74M, but with a starting coordinate of 55268882. When the next alignment is processed and written to the output, the coordinate-sorted order is no longer preserved and the processing crashes with the aforementioned error.  LeftAlignIndels assumes that if the input is coordinate-sorted, the output will be as well. 
    @HD  VN:1.6  SO:coordinate  
    @SQ  SN:chr7  LN:159138663  
    @RG  ID:Ion_V2_BC15_rawlib.basecaller  SM:Ion_V2_BC15_rawlib.basecaller  
    CCAAATAACATGTCTTCT_molbar_1  2064  chr7  55268881  60  56S2M4D2M3I67M  *  0  0  TGATCATCGAATTCTCCAAAATGGCCCGAGACCCCCAGCGCTACCTTGTCATTTAGGGGATGAAAGAATGCATTTGCCAAGTCCTACAGACTCCAACTTCTACCGTGCCCTGATGGATGAAGAAGACATG  <<>>>==>>9?7///)/);;;<7=8>==>>>1>>B>>>;<<<<7<6==<<<7@<8*9988=7+780399///19997=7===8<=<;;=<<<8<9==9??==9====8A===@=9=?@B??<7=;;:;<=  SA:Z:chr7,55268051,-,56M74S,60,*;  AS:i:73  YX:i:41  RG:Z:Ion_V2_BC15_rawlib.basecaller  NM:i:7  MD:Z:2^GGAT69  
    CCAACAGCTGGCAAATGC_molbar_1  2064  chr7  55268881  60  64S24M  *  0  0  CCGTGAGTTGATCATCGAATTCTCCAAAATGGCCCGAGACCCCCAGCGCTACCTTGTCATTCAGGGGGATGAAAGAATGCATTTGCCA  8=====<7<=>>===<=9<7<<<68)77777<6=>===<0;;;;;====>=9=9==>=>9>=<1<<<<<=<7=<<9>>;?89@@<9=9  SA:Z:chr7,55268043,-,64M24S,60,*;  HI:i:1  AS:i:88  RG:Z:Ion_V2_BC15_rawlib.basecaller  NM:i:0  MD:Z:24  
    We are currently using STAR 2.7.9a to generate this alignment, and perhaps a better alignment will be generated with a more recent version of STAR, which we will try next. 

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    Gökalp Çelik

    Hi James Covino

    We were able to replicate the issue and it looks like there is a conflict within the conditional statements within the actual method (Looks like this is a previously uncaught edge case) that checks the read sorting order when reads are fed with coordinate sorted order. Issue does not happen when reads are fed with queryname order therefore the only solution until a fix arrives will be to use queryname sorted bam files to left align indels. Until we have a PR ready we recommend this quick solution for your use case. 

    I hope this helps. 

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