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genomicalignments's Introduction

GenomicAlignments is an R/Bioconductor package for representing and manipulating short genomic alignments (typically aligned reads).

See https://bioconductor.org/packages/GenomicAlignments for more information including how to install the release version of the package (please refrain from installing directly from GitHub).

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genomicalignments's Issues

incomplete use of bamRanges() for summarizeOverlaps method for BamViews

Hello,

I would like to point out, that when I use a BamViews object with the summarizeOverlaps method, the whole Bam file is loaded into Memory, if I do not explicitly provide the param argument.

Here is an example

library(GenomicAlignments)
library(microbenchmark)

tiny_bam <- system.file("extdata", "ex1.bam", package="Rsamtools", mustWork=TRUE)
fl <- c(tiny_bam,tiny_bam)
rngs <- GRanges(c("seq1", "seq2"), IRanges(1, c(15, 15)))
samp <- DataFrame(info=c("ex1","ex2"), row.names=c("ex1","ex2"))
# define the BamViews for multiple files using Rsamtools
view <- BamViews(bamPaths = fl, bamSamples=samp, bamRanges=rngs)

So these function calls will have different memory footprints because in one case we are loading the whole BAM file,

se <- summarizeOverlaps(view, mode=Union, ignore.strand=TRUE)

while in the other we only load the reads that are in the given ranges.

se <- summarizeOverlaps(view, 
                        mode=Union, 
                        ignore.strand=TRUE,
                        param=ScanBamParam(which = rngs))

Best,
Alex

Strand-related warning in summarizeOverlaps() persists in some cases

In release and devel, summarizeOverlaps() produces the following spurious warning:

library(GenomicAlignments)
library(TxDb.Dmelanogaster.UCSC.dm3.ensGene)
exbygene <- exonsBy(TxDb.Dmelanogaster.UCSC.dm3.ensGene, "gene")
library(pasillaBamSubset)
reads <- readGAlignmentsList(untreated3_chr4())
se <- summarizeOverlaps(exbygene, reads)
# Warning message:
# In if (all(strand(reads) == "*")) ignore.strand <- TRUE :
#   the condition has length > 1 and only the first element will be used

Looks like there was an attempt at getting rid of this warning back in 2014 (see 7cfab64) but this fix only addresses the case where reads is a GRangesList object. However reads can be a GAlignments, GAlignmentPairs, GAlignmentsList, GRanges, or GRangesList object, and the fix doesn't address the case where reads is a GAlignmentsList object.

Suggested fix: Replace the first 7 lines (if statement) of internal helper .summarizeOverlaps() with:

    if (!isTRUEorFALSE(ignore.strand))
        stop("'ignore.strand' must be TRUE or FALSE")
    if (!ignore.strand) {
        reads_strand <- strand(reads)
        if (is(reads_strand, "List"))
            reads_strand <- unlist(reads_strand, use.names=FALSE)
        if (all(reads_strand == "*"))
            ignore.strand <- TRUE
    }

The idea is to unlist the strand only if it's a List derivative, which is more robust than unlisting based on the class of reads. The fix also checks ignore.strand which will avoid obscure warnings or errors if the user doesn't supply a TRUE or a FALSE.

Thanks,
H.

> sessionInfo()
R version 3.4.3 (2017-11-30)
Platform: x86_64-pc-linux-gnu (64-bit)
Running under: Ubuntu 16.04.4 LTS

Matrix products: default
BLAS: /home/hpages/R/R-3.4.3/lib/libRblas.so
LAPACK: /home/hpages/R/R-3.4.3/lib/libRlapack.so

locale:
 [1] LC_CTYPE=en_US.UTF-8       LC_NUMERIC=C              
 [3] LC_TIME=en_US.UTF-8        LC_COLLATE=en_US.UTF-8    
 [5] LC_MONETARY=en_US.UTF-8    LC_MESSAGES=en_US.UTF-8   
 [7] LC_PAPER=en_US.UTF-8       LC_NAME=C                 
 [9] LC_ADDRESS=C               LC_TELEPHONE=C            
[11] LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C       

attached base packages:
[1] stats4    parallel  stats     graphics  grDevices utils     datasets 
[8] methods   base     

other attached packages:
 [1] pasillaBamSubset_0.16.0                  
 [2] TxDb.Dmelanogaster.UCSC.dm3.ensGene_3.2.2
 [3] GenomicFeatures_1.30.3                   
 [4] AnnotationDbi_1.40.0                     
 [5] GenomicAlignments_1.14.1                 
 [6] Rsamtools_1.30.0                         
 [7] Biostrings_2.46.0                        
 [8] XVector_0.18.0                           
 [9] SummarizedExperiment_1.8.1               
[10] DelayedArray_0.4.1                       
[11] matrixStats_0.53.1                       
[12] Biobase_2.38.0                           
[13] GenomicRanges_1.30.3                     
[14] GenomeInfoDb_1.14.0                      
[15] IRanges_2.12.0                           
[16] S4Vectors_0.16.0                         
[17] BiocGenerics_0.24.0                      

loaded via a namespace (and not attached):
 [1] Rcpp_0.12.16           compiler_3.4.3         prettyunits_1.0.2     
 [4] bitops_1.0-6           tools_3.4.3            zlibbioc_1.24.0       
 [7] progress_1.1.2         biomaRt_2.34.2         digest_0.6.15         
[10] bit_1.1-12             RSQLite_2.0            memoise_1.1.0         
[13] lattice_0.20-35        pkgconfig_2.0.1        Matrix_1.2-12         
[16] DBI_0.8                GenomeInfoDbData_1.0.0 rtracklayer_1.38.3    
[19] httr_1.3.1             stringr_1.3.0          bit64_0.9-7           
[22] grid_3.4.3             R6_2.2.2               XML_3.98-1.10         
[25] RMySQL_0.10.14         BiocParallel_1.12.0    magrittr_1.5          
[28] blob_1.1.1             assertthat_0.2.0       stringi_1.1.7         
[31] RCurl_1.95-4.10       

mapFromAlignments() and soft-clipping

This looks wrong:

library(GenomicAlignments)
mypos <- IRanges(1:7, width=1, names=rep("read1", 7))
alignment <- GAlignments("chr1", pos=1L, cigar="2S3M2D7M", strand("+"), names="read1")
mapFromAlignments(mypos, alignment)
# IRanges object with 7 ranges and 0 metadata columns:
#             start       end     width
#         <integer> <integer> <integer>
#   read1         1         1         1
#   read1         2         2         1
#   read1         3         3         1
#   read1         4         4         1
#   read1         5         5         1
#   read1         8         8         1
#   read1         9         9         1

The reported deletion (ref pos 6 & 7) seems wrong. Should be ref pos 4 & 5.

sessionInfo():

> sessionInfo()
R Under development (unstable) (2019-10-30 r77336)
Platform: x86_64-pc-linux-gnu (64-bit)
Running under: Ubuntu 16.04.6 LTS

Matrix products: default
BLAS:   /home/hpages/R/R-4.0.r77336/lib/libRblas.so
LAPACK: /home/hpages/R/R-4.0.r77336/lib/libRlapack.so

locale:
 [1] LC_CTYPE=en_US.UTF-8       LC_NUMERIC=C              
 [3] LC_TIME=en_US.UTF-8        LC_COLLATE=en_US.UTF-8    
 [5] LC_MONETARY=en_US.UTF-8    LC_MESSAGES=en_US.UTF-8   
 [7] LC_PAPER=en_US.UTF-8       LC_NAME=C                 
 [9] LC_ADDRESS=C               LC_TELEPHONE=C            
[11] LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C       

attached base packages:
[1] stats4    parallel  stats     graphics  grDevices utils     datasets 
[8] methods   base     

other attached packages:
 [1] GenomicAlignments_1.23.1    Rsamtools_2.3.4            
 [3] Biostrings_2.55.4           XVector_0.27.0             
 [5] SummarizedExperiment_1.17.2 DelayedArray_0.13.4        
 [7] BiocParallel_1.21.2         matrixStats_0.55.0         
 [9] Biobase_2.47.2              GenomicRanges_1.39.2       
[11] GenomeInfoDb_1.23.13        IRanges_2.21.3             
[13] S4Vectors_0.25.12           BiocGenerics_0.33.0        

loaded via a namespace (and not attached):
 [1] lattice_0.20-38        crayon_1.3.4           bitops_1.0-6          
 [4] grid_4.0.0             zlibbioc_1.33.1        Matrix_1.2-18         
 [7] tools_4.0.0            RCurl_1.98-1.1         compiler_4.0.0        
[10] GenomeInfoDbData_1.2.2

Permission for SweaveToRmd

Hi,
I'm volunteering with Sweave2Rmd and we're trying to replace all Bioconductor Sweave vignettes R Markdown
vignettes. Would you be willing to accept a pull request with the R Markdown
version of GenomicAlignments? I also wanted to ask if you can assign this to me on the board?
Thanks for your time.

Error in isCompatibleWithSkippedExons() for single-end read alignments w/ more than 3 junctions

Hi, single-end read alignments with more than 3 junctions in the cigar string cause the following error:

suppressPackageStartupMessages(library(GenomicAlignments))
suppressPackageStartupMessages(library(TxDb.Hsapiens.UCSC.hg38.knownGene))

txdb <- TxDb.Hsapiens.UCSC.hg38.knownGene
tx <- exonsBy(txdb, by="tx")
aln <- GAlignments("chr1", pos=42837180L, cigar="5S11M1710N21M1111N21M98N27M91N16M",
                   strand("+"), names="read1")
ovtx <- findOverlaps(GRanges(aln), tx, ignore.strand=FALSE)
ovtxenc <- encodeOverlaps(grglist(aln), tx, hits=ovtx,
                          flip.query.if.wrong.strand=FALSE)
maskskpovtx <- isCompatibleWithSkippedExons(ovtxenc) # error
Error in paste0(build_compatible_encoding_subpatterns(njunc), collapse = skipped_exons_subpatterns[njunc +  : 
  invalid 'collapse' argument

Here is my session information:

sessionInfo()
R version 4.3.2 (2023-10-31)
Platform: x86_64-apple-darwin20 (64-bit)
Running under: macOS Ventura 13.6.1

Matrix products: default
BLAS:   /Library/Frameworks/R.framework/Versions/4.3-x86_64/Resources/lib/libRblas.0.dylib 
LAPACK: /Library/Frameworks/R.framework/Versions/4.3-x86_64/Resources/lib/libRlapack.dylib;  LAPACK version 3.11.0

locale:
[1] en_US.UTF-8/en_US.UTF-8/en_US.UTF-8/C/en_US.UTF-8/en_US.UTF-8

time zone: Europe/Madrid
tzcode source: internal

attached base packages:
[1] stats4    stats     graphics  grDevices utils     datasets  methods   base     

other attached packages:
 [1] TxDb.Hsapiens.UCSC.hg38.knownGene_3.18.0 GenomicFeatures_1.54.1                  
 [3] AnnotationDbi_1.64.1                     GenomicAlignments_1.38.0                
 [5] Rsamtools_2.18.0                         Biostrings_2.70.1                       
 [7] XVector_0.42.0                           SummarizedExperiment_1.32.0             
 [9] Biobase_2.62.0                           MatrixGenerics_1.14.0                   
[11] matrixStats_1.1.0                        GenomicRanges_1.54.1                    
[13] GenomeInfoDb_1.38.1                      IRanges_2.36.0                          
[15] S4Vectors_0.40.2                         BiocGenerics_0.48.1                     
[17] colorout_1.3-0                          

loaded via a namespace (and not attached):
 [1] KEGGREST_1.42.0         rjson_0.2.21            lattice_0.22-5          vctrs_0.6.5            
 [5] tools_4.3.2             bitops_1.0-7            generics_0.1.3          curl_5.2.0             
 [9] parallel_4.3.2          tibble_3.2.1            fansi_1.0.6             RSQLite_2.3.4          
[13] blob_1.2.4              pkgconfig_2.0.3         Matrix_1.6-4            dbplyr_2.4.0           
[17] lifecycle_1.0.4         GenomeInfoDbData_1.2.11 compiler_4.3.2          stringr_1.5.1          
[21] progress_1.2.3          codetools_0.2-19        yaml_2.3.7              RCurl_1.98-1.13        
[25] pillar_1.9.0            crayon_1.5.2            BiocParallel_1.36.0     DelayedArray_0.28.0    
[29] cachem_1.0.8            abind_1.4-5             tidyselect_1.2.0        digest_0.6.33          
[33] stringi_1.8.2           restfulr_0.0.15         dplyr_1.1.4             biomaRt_2.58.0         
[37] fastmap_1.1.1           grid_4.3.2              cli_3.6.2               SparseArray_1.2.2      
[41] magrittr_2.0.3          S4Arrays_1.2.0          XML_3.99-0.16           utf8_1.2.4             
[45] rappdirs_0.3.3          filelock_1.0.3          prettyunits_1.2.0       bit64_4.0.5            
[49] httr_1.4.7              bit_4.0.5               png_0.1-8               hms_1.1.3              
[53] memoise_2.0.1           BiocIO_1.12.0           BiocFileCache_2.10.1    rtracklayer_1.62.0     
[57] rlang_1.1.2             glue_1.6.2              DBI_1.1.3               xml2_1.3.6             
[61] R6_2.5.1                zlibbioc_1.48.0        

I've submitted a PR that fixes it.

cheers,

robert.

Differences in reads' base content with IGV.

Hey there and thanks for the amazing package, it's been really helpful!

I stumbled upon an issue which I would like to share with you.
So I have generated a synthetic bam file using NEAT that contains reads in the TP53 region (it's located in the 17th chromosome and spans for about 19k bases).
My interest is to benchmark the somatic variants which are produced by GATK-Mutect2 and to do that I need to know exactly the Allele Frequencies (AF) of the variants and hence the number of all bases in each chromosomal position.

To my great surprise I found that at certain chromosomal positions, although the coverage is correct, the reported bases do differ!
Below I present the lines of code that I am using in R with the help of "GenomicAlignments" library.

library(GenomicAlignments)

bam = scanBam("1/1_golden.bam")

#Read genomic alignments
olaps <- readGAlignments("1/1_golden.bam")

#chromosomal position
p=474

# reads in chromosomal position
index = which(start(olaps) <= p & end(olaps) >= p)

#extract reads from bam
my_seqs = bam[[1]]$seq[index]

#distance from position p
ret = p - start(olaps[index, ]) + 1

out = mapply(function(x, y) {
    
    str_sub(x, y, y)
    
}, as.character(my_seqs), ret)


out = data.table(
    "base" = out
)

out = out[, by = base, .N] 

I am also attaching a print screen.
IGV

On the left you see the out parameter which contains the bases from the chromosomal position p (in this case p=474) and on the right you see the info as produced by IGV for the same position p. The coverage is the same (478 in both cases), the difference is found in the base content of the reads.

I am very curious to see what causes this systematic deviation between IGV and my code. If anyone has faced this problem before or has any clue, I would really appreciate it!

Thanks in advance,
Stella

S4Vectors

sorry, I have a question,

when I installed the package in R 4.0.2, it was wrong with the S4Vectors 0.26.1 but need to >=0.27.12, how to solve this question?

Thanks

reference to query location

Does the opposite function exist to queryLocs2refLocs ?

i.e. given a reference location, finding the position in the query that corresponds to it. This would be a basic useful function.

Thanks.

Clarify Strandedness Options for SummarizeOverlaps in Documentation

Hey there,

It seems to me that due to how most RNAseq libraries are now constructed, most people who want to count reads per feature with summarizeOverlaps will actually want to use ignore.strand = FALSE and preprocess.reads = invertStrand. However, I only stumbled across this after realizing that my output was wildly different from what htseq-count with -s reverse gave me when I only used ignore.strand = FALSE.

That you need to use preprocess.reads = invertStrand with most modern stranded RNAseq libraries is not mentioned in the documentation for summarizeOverlaps or in the example RNAseq workflow (which is not this package, but it's still confusing). The only information that I could find to lead me to this was in the bioconductor support forums.

Could the documentation be clarified in some way? I could see this tripping up new users.

summarizeJunctions with genome fails if no junction exists

This is the example code to reproduce the error:

library(GenomicAlignments)
library(BSgenome.Hsapiens.UCSC.hg19)

aln1_file <- system.file("extdata", "ex1.bam", package="Rsamtools")
aln1 <- readGAlignments(aln1_file)

summarizeJunctions(aln1, genome="BSgenome.Hsapiens.UCSC.hg19")

This will produces the following error message:

Error in .get_xscat_ans_cardinality(...) : 
  xscat() cannot mix arguments made of 0 sequence with arguments made of 1 or more sequences
In addition: Warning message:
In .Seqinfo.mergexy(x, y) :
  The 2 combined objects have no sequence levels in common. (Use
  suppressWarnings() to suppress this warning.)

Error using granges with unused argument on.discordant.seqnames

Dear Sir,
I have used granges function in this package for a long time. However, I got an error after updating GenomicAlignments (version 1.16.0) through bioconductor.
I used granges like below

t.record.range <- granges(temp, on.discordant.seqnames="drop");
Error in .local(x, use.names, use.mcols, ...) : 
  unused argument (on.discordant.seqnames = "drop")

the argument on.discordant.seqnames seemed to be removed.
I have search the solution using google, but I got nothing.
Now my question are:

  1. If I simply remove on.discordant.seqnames='drop' in my code, is the effect the same? Or could you please point out the solution?
  2. The document in bioconductor is different with the help in R (type ?granges)
    https://bioconductor.org/packages/release/bioc/manuals/GenomicAlignments/man/GenomicAlignments.pdf

Non-graceful, when trying to access metadata column from GAlignment object

Hi,

by accident I tried to access the metadata column from GAlignment object directly via $. This ended up in a bit ungraceful exit.

library(GenomicAlignments)
ga <- readGAlignments(file, param = ScanBamParam(what = "qual"))
ga$qual # this crashes
#> Error: evaluation nested too deeply: infinite recursion / options(expressions=)?
mcols(ga)$qual # this works
#>   A PhredQuality instance of length x
#>          width seq

I don't know, if that is expected behavior or it is just the way it needs to be (please close the issue if that is the case).

However, a more graceful exit might benefit understanding the GAlignment class. I stumbled onto this by assuming it might work like a GRanges/DataFrame object.

Felix

sessionInfo()
#> R version 3.6.0 (2019-04-26)
#> Platform: x86_64-w64-mingw32/x64 (64-bit)
#> Running under: Windows 10 x64 (build 18362)
#> 
#> Matrix products: default
#> 
#> locale:
#> [1] LC_COLLATE=German_Germany.1252  LC_CTYPE=German_Germany.1252   
#> [3] LC_MONETARY=German_Germany.1252 LC_NUMERIC=C                   
#> [5] LC_TIME=German_Germany.1252    
#> 
#> attached base packages:
#> [1] stats4    parallel  stats     graphics  grDevices utils     datasets 
#> [8] methods   base     
#> 
#> other attached packages:
#>  [1] RNAmodR.Data_0.99.5         ExperimentHubData_1.10.0   
#>  [3] AnnotationHubData_1.14.0    ExperimentHub_1.10.0       
#>  [5] AnnotationHub_2.17.0        BiocFileCache_1.8.0        
#>  [7] dbplyr_1.4.1                futile.logger_1.4.3        
#>  [9] GenomicAlignments_1.20.0    Rsamtools_2.0.0            
#> [11] Biostrings_2.52.0           XVector_0.24.0             
#> [13] SummarizedExperiment_1.14.0 DelayedArray_0.10.0        
#> [15] BiocParallel_1.17.18        matrixStats_0.54.0         
#> [17] Biobase_2.44.0              GenomicRanges_1.36.0       
#> [19] GenomeInfoDb_1.20.0         IRanges_2.18.1             
#> [21] S4Vectors_0.22.0            BiocGenerics_0.30.0        
#> 
#> loaded via a namespace (and not attached):
#>  [1] bitops_1.0-6                  bit64_0.9-7                  
#>  [3] progress_1.2.2                httr_1.4.0                   
#>  [5] tools_3.6.0                   R6_2.4.0                     
#>  [7] DBI_1.0.0                     tidyselect_0.2.5             
#>  [9] prettyunits_1.0.2             bit_1.1-14                   
#> [11] curl_3.3                      compiler_3.6.0               
#> [13] graph_1.62.0                  BiocCheck_1.20.0             
#> [15] formatR_1.7                   rtracklayer_1.44.0           
#> [17] RBGL_1.60.0                   rappdirs_0.3.1               
#> [19] stringr_1.4.0                 digest_0.6.19                
#> [21] rmarkdown_1.13                stringdist_0.9.5.2           
#> [23] AnnotationForge_1.26.0        rBiopaxParser_2.24.0         
#> [25] pkgconfig_2.0.2               htmltools_0.3.6              
#> [27] highr_0.8                     rlang_0.3.4                  
#> [29] RSQLite_2.1.1                 shiny_1.3.2                  
#> [31] jsonlite_1.6                  dplyr_0.8.1                  
#> [33] RCurl_1.95-4.12               magrittr_1.5                 
#> [35] GenomeInfoDbData_1.2.1        Matrix_1.2-17                
#> [37] Rcpp_1.0.1                    stringi_1.4.3                
#> [39] yaml_2.2.0                    zlibbioc_1.30.0              
#> [41] biocViews_1.52.2              grid_3.6.0                   
#> [43] blob_1.1.1                    promises_1.0.1               
#> [45] crayon_1.3.4                  lattice_0.20-38              
#> [47] GenomicFeatures_1.36.1        hms_0.4.2                    
#> [49] knitr_1.23                    pillar_1.4.1                 
#> [51] optparse_1.6.2                RUnit_0.4.32                 
#> [53] codetools_0.2-16              biomaRt_2.40.0               
#> [55] futile.options_1.0.1          XML_3.98-1.20                
#> [57] glue_1.3.1                    evaluate_0.14                
#> [59] lambda.r_1.2.3                data.table_1.12.2            
#> [61] BiocManager_1.30.4            httpuv_1.5.1                 
#> [63] getopt_1.20.3                 purrr_0.3.2                  
#> [65] assertthat_0.2.1              xfun_0.7                     
#> [67] mime_0.6                      xtable_1.8-4                 
#> [69] later_0.8.0                   tibble_2.1.3                 
#> [71] OrganismDbi_1.26.0            AnnotationDbi_1.46.0         
#> [73] memoise_1.1.0                 interactiveDisplayBase_1.22.0

cigarNarrow_segmentationFault

PACKAGE VERSION

GenomicAlignments (1.32.0)

PROBLEM

I have trouble to subset a cigar string for a region larger than 100,000 in length.
Is this a memory issue on my PC or a know limitation of cigarNarrow function?

INPUT CIGAR

cg <- "7=1X4=1X2=1X19=2X3=2X2=1X6=1X14=1X3=1X18=1X39=1X7=1D90=1X3=1X13=1X4=1X13=6I97=1X213=4D24=1X50=1X151=18I166=1X16=1X4=1X46=3D58=1X144=1X182=1X13=1X127=1X4=1X35=1X66=1X141=1X119=1X42=1X35=6I126=1X23=1X12=1X35=1X9=1X12=1X71=255D8=1X57=2X24=1X22=1X3=1X54=1X4=1X22=1X30=1X66=2X107=1X3=1X112=1X257=1X7=1X21=1X126=1X55=1X174=3D137=1X186=1X54=1X30=1X42=1X55=1X40=2I2X133=1X59=1X73=1D103=1X8=1X101=1X22=1X71=1X56=1X35=1X33=2X14=1X185=1X32=1X47=1X11=4I5=3X33=1X38=1X69=1X60=4I30=1X9=1X34=2X100=1X1=1X68=1X14=1X7=1X53=1X25=5I1X52=5I7=1X302=1X240=1X105=1X41=1X29=1X34=1X27=3D246=1X56=1X210=1X144=1X116=1X166=1X59=1X1=1X13=5I66=1X107=1X48=1X8=2X12=1X6=1X45=1X93=1X43=1X11=1X73=1X27=1X26=1X26=1X7=1X9=1D13=1X49=1X106=2D1=1X147=1X81=1X51=1X78=1X36=9523D11=1X6=1X23=1X108=1X49=1X7=2X11=1I179=1D62=1X85=1X27=1X21=1X11=1X2=1X208=1X16=1X24=1X2=1X9=1X39=1X90=1I13=5I84=1X46=1X93=1X124=1X7=1X22=10I24=1X35=1X53=1X80=1X6=1X67=1X73=1I3=1X3=1X8=11I91=1X242=2X51=1X165=1X103=1X21=1X152=1I34=1X153=1X29=1X20=1X25=1X71=1X12=1X117=2D32=1X72=1X93=1X1=1X3=1X34=1X49=1X10=1X17=1X39=1X26=1X15=1X35=9I28=1X6=1X43=1X57=1X10=1X190=1X149=1X52=1X32=1X1=1X59=2X50=1X339=1X24=1X42=1I11=5I9=1X584=1X216=1X15=1X11=1X60=1X5=1X245=1X140=1X15=1I204=1X448=1X26=1X24=1X35=1X37=1X16=1X52=1X10=1X93=1X87=1X91=1X53=1X5=1X24=1X115=1X74=1X249=1X25=1I65=1X145=1X177=1X11=1X1=1X20=1I1X107=1X47=1X9=1X2=2D21=1D101=1X48=1X119=1X24=1X78=1X61=1X1=1X4=1X2=1X6=1X1=1X10=2X8=2X2=2I1X1=1X5=1X2=1X3=1X4=1X106=1I161=1D109=1X68=1X2=1X384=1X64=1X145=1X38=1X49=4D31=1X205=2X17=1X19=1X2=1X72=1X86=1X71=5I264=1X13=2X150=1X33=1X253=1X29=1X159=1X28=1X19=1X2=1X7=1X42=1X24=1X25=1D15=6I62=1X86=1X155=1X44=1X5=1X277=1X48=1X224=1X248=1X358=1X65=1X98=1X107=1X216=1X23=1X120=1X41=1X54=1D60=1X55=1X27=1X17=2D1=1X12=1X30=1X1=1X2=1X5=1D6=1X12=1X4=2D36=12I5=1X7=1X12=1X6=1X3=1X7=12I212=4D73=1X14=1X39=1X96=1X99=2X169=1X53=1X237=1X37=1X14=1X63=1X222=1X32=1X12=2X80=1X4=10I20=1X120=1X68=1X18=1X50=1X2=4D256=1X421=1X91=3I67=1X6=1X28=1X14=1X3=1X45=1X38=1X19=2D189=3D92=2X43=1X74=1X92=1X165=1X155=5D1X15=1X1=1X67=1X101=1X24=1X61=1X58=1I187=1X1=3D7=2X241=1X24=1X70=1X69=1X25=1X143=6I15=1X96=1X1=1X45=1X27=1X19=1X29=1X46=1X188=1X154=1X78=1X84=1I13=1X78=1X39=1X343=1X2=1X61=1X143=1X20=1X41=1X30=1X16=25I15=1X16=1X1=1X44=1X43=1X17=8I183=9D113=1X41=1X42=1X7=1X2=1X7=1X133=1X17=1X63=1X42=1X1=3I7X21=1X92=1X191=1X86=5I129=1X8=1X25=1I34=1X137=1X21=4D131=1X127=1X68=1X238=1X135=1X36=1X16=1X83=1X7=1I8=2X93=1X13=1X20=1X68=1D4=1X52=1X20=1X40=1X44=1X1=1X41=5I48=1X112=1X33=1X81=1X18=1X33=1X50=1X47=1X87=5I10=1X7=13I13=1X4=1X64=1X75=1X92=1X11=2X137=1X108=1X12=1D19=1X112=3D32=1X40=1X81=5I59=1X21=1X60=1I19=1X26=1X67=1X44=4D56=1X90=1X2=8D243=1X28=1X150=1X392=1X27=2D64=1X453=2D14=1X49=1X119=1X27=1I2=1X542=1X78=1I50=1X49=1X86=1X53=1X18=1X14=1X1=1X4=1X25=1X18=2I175=1X37=4D57=1X105=6D176=1X55=1X184=1X1=1X40=1X91=1X40=1X28=1X13=1X66=1X2=1X72=1X112=1X63=1X207=1X65=1X157=1X362=1X8=1X16=1X29=1X16=1X6=1X290=2X15=1X34=1X286=1X224=1X111=1X66=1X12=1X59=1X80=1X4=1I62=1X153=1X138=1X40=1X77=1X184=1X140=5D74=1X38=1X32=1X62=1X20=1X24=1X110=4D36=1X44=1X79=1X78=1X246=1X45=1X17=1X62=1X79=1X146=1X41=8I1=1X100=1X156=1X79=1X121=1X80=1X23=1X24=1X339=1X190=1X69=1X216=25I73=1X186=1X54=1X98=1X152=1X7=1X71=1X29=1X9=1X9=15D109=1X30=5D111=1X48=1X20=4I104=4I243=1X14=1X55=1X152=1X138=1X106=1I117=1X152=1X15=1X100=1X7=1X5=12D188=1X14=1I28=1X36=1X106=1X187=1X37=1X6=1X3=2X3=6I29=1X31=1X39=1X42=1X50=1X104=2X14=1X29=1X16=1X76=3X8=14I109=1X20=1X48=1X60=1X9=1X37=2X76=1X7=1X58=1X186=1X77=1X83=1X91=1X44=1X84=1X27=1X212=1X80=1X6=1X78=1X3=1X65=1X5=1I158=1X64=1X46=1X62=1X7=2D22=1X41=1X3=1X71=1X13=1X2=1X23=1X27=1X30=1X27=1X4=1X69=1I2=1X32=1D19=1X109=1X19=1X25=1X368=5I13=1X120=1X44=1X54=1X66=9I2=1X100=1X94=7D94=1X6=1X10=1X13=1X64=1X21=1X33=1X4=1X69=1X79=1X349=1X150=1X18=1X121=1X87=1X6=1X1=1X63=3D209=1I8=1X7=1X93=1X38=1X78=1X48=1X15=1D53=1X14=1X129=1X110=1X18=5I64=1X31=2X94=1X26=1X3=1X6=1X58=1X3=1X4=2X31=1X152=1X111=1X63=1X144=1X32=11D6=1X11=1X59=1X23=1D6=1X212=1X91=1X8=1X14=1X31=1X10=1X94=1X5=1X28=3D3=1X3=1X135=142D2=1X17=1I119=1X123=6D160=1X163=1X19=10I65=1X53=1X1040=1X98=1X38=1X253=1X233=1X17=1D13=11D9=1X63=1X23=1X21=1X306=1X7=1X9=1X1=1X57=1X13=1D95=1I1X55=1X220=1X35=1X277=1X52=2D2=1X44=1X13=3D257=1X4=1X72=1X117=1X162=1X37=1X130=1X5=1X45=1X182=25D80=1I41=5D45=1X49=1X103=1X75=1X6=1X88=1X99=1X76=1I2=1X8=1D7=2I2=1X4=2I186=1X147=1X75=1X44=1X56=1X80=1X22=1I51=1X28=1X4=2D24=2X70=2X4=1D4=1X36=1X36=1I122=1X12=1X31=1X36=1X66=2I155=1X241=3I24=1X359=1X96=1X72=1X467=1I168=8I143=1X27=1I412=1X219=1X134=1X424=1X225=1X486=1X258=1X2=1X50=1X88=1X125=1X70=4I1222=1X1342=1X554=1X22=1X34=1D570=1D64=1X119=1X213=1X1=1X20=1X66=1X22=1X905=1X25=1X221=3D518=1D86=1X1135=1X729=2D80=1X132=1X24=1I213=1X63=1X7=1X80=1X77=1X53=3D13=1X125=1X13=1X39=1X158=1X16=1X15=1X53=3D5=2X37=1X51=1X57=1X11=1X10=3D210=2X396=1X227=1X102=1X6=1X166=1X110=1X19=1X51=2X16=1X134=1X132=1X46=1X197=1X194=1X60=3I193=1X130=2X60=1X110=1X9=1X56=1X18=1X167=1X62=1X11=3D52=1X8=1X39=1X29=1X125=1X27=1X43=1X65=1X9=1X213=2I53=1X12=2D116=1X62=1X32=1X213=1X43=1X98=1X35=1D57=1X122=1X75=1X45=2X21=2I12=1X2=1X18=1X54=1X19=1X35=1X17=1X48=1X13=1X4=3D31=1X8=1X75=1X17=1X200=1X11=1X9=1X83=1X70=1D92=1X14=1X31=1X28=1X86=1X154=1X3=1X30=1X22=1X130=1X13=1X1=1I129=1X12=2X130=1X13=1X9=1X8=1X11=1X63=5D7=1X261=1X290=1X9=1X1=1X173=7D30=1X10=1X121=1X58=1X42=7I9=3X25=1X24=1X100=1X27=1X30=3I171=2I52=1X31=1X155=1X16=1X18=1X39=2X4=1X637=1X5=1X60=1X49=4I40=1X11=1X204=1X30=1X98=1X8=1X43=1X151=1D141=1X14=1X95=1X14=1X83=1X647=1X69=1X53=1X285=1X418=1I29=1X181=1X41=1X44=1X89=1X104=1X525=1X78=1X180=5I159=1X2=1X5=621D14=1X8=1X30=1X2=1X5=2X112=1I191=1X287=1X46=6D88=1X3=1X107=1X54=1X872=1X461=1X1521=1X666=1X226=1X89=1X66=1X211=1X4=1X1290=1X299=1I158=1X39=1X197=1X113=22D79=1X18=15I48=1X78=1X157=16D3=2D18=1X124=4I11=1X408=1X265=2D120=1X18=1X18=1X49=1X156=1X115=1X6=1X4=1X55=1X12=1D103=1X2=2I41=1X235=1X7=1X106=1X111=1X3=1X112=1X7=1X169=10I1=1X99=1X140=1X5=1X100=1X3=1X579=6D90=1X41=22D77=1X98=1X2=1X61=1X69=1I249=1X34=1X1=1X17=1X94=2D8=1X118=1X29=1X133=2X341=1X28=2D248=1X25=1X106=1X67=1X115=1X133=1X41=1X135=1X444=1X153=3D53=1X104=4I2=1X15=1X331=1D176=2I202=1X47=1X27=2X41=1X9=1D114=1X60=14I8=1X28=2I20=2D147=1X153=1X205=1X40=1X40=1X16=5D154=1X49=3D13=1X581=1X612=4I49=1X1=28I263=2D50=1X57=1X38=1X327=1X65=1X515=1X98=1X3=4D126=1X18=1X81=1X10=1I88=1X156=1X30=1X82=1X4=1X86=1D271=1X53=1X38=1X67=1X11=4D115=1X59=1X2=1X175=1X195=1X107=1X90=1X166=1I86=1X154=1X411=1X136=1X33=1X75=1X114=1X110=1X413=1X58=1X335=1X266=1X88=6I32=1X1521=1X701=1X186=9D257=1X154=1X106=1X41=1I387=1X79=1X91=2D2007=12D226=1X58=1X262=1X1585=1X63=1X264=1X301=1X180=1X106=1X558=1X178=1X44=3D84=1X25=1X20=1X8=1X212=2X148=1I88=1X168=1X8=1X535=2I51=3D1X421=1X124=1X1=1X32=1X94=1X110=3D33=1X12=1X17=1X291=1X433=1X1596=1X111=1X212=1X122=1X55=1D81=1X326=1X137=1X261=1I131="

RUN COMMAND

GenomicAlignments::cigarNarrow(cigar = cg, start = 8703, end = 124563)[1]

ERROR message

*** caught segfault ***
address 0x7f879f4ed000, cause 'invalid permissions'

Traceback:
1: .Call2("cigar_narrow", cigar, width(threeranges$left), width(threeranges$right), PACKAGE = "GenomicAlignments")
2: GenomicAlignments::cigarNarrow(cigar = paf.aln$cg, start = 8703, end = 124563)

Possible actions:
1: abort (with core dump, if enabled)
2: normal R exit
3: exit R without saving workspace
4: exit R saving workspace
Selection:

SessionInfo

R version 4.1.1 (2021-08-10)
Platform: x86_64-pc-linux-gnu (64-bit)
Running under: Ubuntu 20.04.3 LTS

Matrix products: default
BLAS: /usr/lib/x86_64-linux-gnu/blas/libblas.so.3.9.0
LAPACK: /usr/lib/x86_64-linux-gnu/lapack/liblapack.so.3.9.0

locale:
[1] LC_CTYPE=en_US.UTF-8 LC_NUMERIC=C LC_TIME=en_US.UTF-8 LC_COLLATE=en_US.UTF-8
[5] LC_MONETARY=en_US.UTF-8 LC_MESSAGES=en_US.UTF-8 LC_PAPER=en_US.UTF-8 LC_NAME=C
[9] LC_ADDRESS=C LC_TELEPHONE=C LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C

attached base packages:
[1] stats graphics grDevices utils datasets methods base

loaded via a namespace (and not attached):
[1] rstudioapi_0.13 XVector_0.34.0 GenomicRanges_1.46.1 BiocGenerics_0.40.0
[5] zlibbioc_1.40.0 GenomicAlignments_1.32.0 IRanges_2.28.0 BiocParallel_1.28.3
[9] lattice_0.20-45 GenomeInfoDb_1.30.1 tools_4.1.1 SummarizedExperiment_1.24.0
[13] parallel_4.1.1 grid_4.1.1 Biobase_2.54.0 matrixStats_0.62.0
[17] crayon_1.5.1 Matrix_1.3-4 GenomeInfoDbData_1.2.7 S4Vectors_0.32.4
[21] bitops_1.0-7 RCurl_1.98-1.7 DelayedArray_0.20.0 compiler_4.1.1
[25] MatrixGenerics_1.6.0 Biostrings_2.62.0 Rsamtools_2.10.0 stats4_4.1.1

readGAlignmentPairs causes Floating point exception (core dumped)

hi there,

I am getting a core dump trying to use readGAlignmentPairs to read in a bam file (actually a set of bam files). I can use readGAlignments on the same bam file without any trouble.

I get a similar issue whether I'm running R on a node of our linux cluster or on my mac. On the linux cluster it seems to happen every time I try to read the file, and on the mac it's a bit more hit and miss - sometimes it works fine, but repeating the same command a few seconds later causes R to crash. Weird.

Some code is below. I think I'm using the latest release versions of everything. I'm happy to upload a bam file somewhere if that's useful.

thanks!

Janet

on linux:

library(GenomicAlignments)
sessionInfo()

R version 3.6.2 (2019-12-12)
Platform: x86_64-pc-linux-gnu (64-bit)
Running under: Ubuntu 14.04.5 LTS

Matrix products: default
BLAS/LAPACK: /app/easybuild/software/OpenBLAS/0.2.18-GCC-5.4.0-2.26-LAPACK-3.6.1/lib/libopenblas_prescottp-r0.2.18.so

locale:
 [1] LC_CTYPE=en_US.UTF-8       LC_NUMERIC=C              
 [3] LC_TIME=en_US.UTF-8        LC_COLLATE=en_US.UTF-8    
 [5] LC_MONETARY=en_US.UTF-8    LC_MESSAGES=en_US.UTF-8   
 [7] LC_PAPER=en_US.UTF-8       LC_NAME=C                 
 [9] LC_ADDRESS=C               LC_TELEPHONE=C            
[11] LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C       

attached base packages:
[1] stats4    parallel  stats     graphics  grDevices utils     datasets 
[8] methods   base     

other attached packages:
 [1] GenomicAlignments_1.22.1    Rsamtools_2.2.1            
 [3] Biostrings_2.54.0           XVector_0.26.0             
 [5] SummarizedExperiment_1.16.1 DelayedArray_0.12.2        
 [7] BiocParallel_1.20.1         matrixStats_0.55.0         
 [9] Biobase_2.46.0              GenomicRanges_1.38.0       
[11] GenomeInfoDb_1.22.0         IRanges_2.20.2             
[13] S4Vectors_0.24.3            BiocGenerics_0.32.0        

loaded via a namespace (and not attached):
[1] lattice_0.20-38        bitops_1.0-6           grid_3.6.2            
[4] zlibbioc_1.32.0        Matrix_1.2-18          tools_3.6.2           
[7] RCurl_1.95-4.12        compiler_3.6.2         GenomeInfoDbData_1.2.2

temp1 <- readGAlignments("mapToCombinedFlyYeastHumanGenomes_ATfiltReads/just_dm6_reads/zz_testR/Nnk_chip_rep3.ATfilt.combined.dm6.chr4.bam")

temp1
# GAlignments object with 421653 alignments and 0 metadata columns:
#            seqnames strand       cigar    qwidth     start       end     width
#               <Rle>  <Rle> <character> <integer> <integer> <integer> <integer>
#        [1]     chr4      -        151M       151         7       157       151
#        [2]     chr4      +      54M97H        54        24        77        54
## etc 

temp2 <- readGAlignmentPairs("mapToCombinedFlyYeastHumanGenomes_ATfiltReads/just_dm6_reads/zz_testR/Nnk_chip_rep3.ATfilt.combined.dm6.chr4.bam")
## Floating point exception (core dumped)

and on the mac

library(GenomicAlignments)
sessionInfo()

R version 3.6.2 (2019-12-12)
Platform: x86_64-apple-darwin15.6.0 (64-bit)
Running under: macOS Mojave 10.14.6

Matrix products: default
BLAS:   /Library/Frameworks/R.framework/Versions/3.6/Resources/lib/libRblas.0.dylib
LAPACK: /Library/Frameworks/R.framework/Versions/3.6/Resources/lib/libRlapack.dylib

locale:
[1] en_US.UTF-8/en_US.UTF-8/en_US.UTF-8/C/en_US.UTF-8/en_US.UTF-8

attached base packages:
[1] stats4    parallel  stats     graphics  grDevices utils     datasets  methods   base     

other attached packages:
 [1] GenomicAlignments_1.22.1    Rsamtools_2.2.1             Biostrings_2.54.0          
 [4] XVector_0.26.0              SummarizedExperiment_1.16.1 DelayedArray_0.12.2        
 [7] BiocParallel_1.20.1         matrixStats_0.55.0          Biobase_2.46.0             
[10] GenomicRanges_1.38.0        GenomeInfoDb_1.22.0         IRanges_2.20.2             
[13] S4Vectors_0.24.3            BiocGenerics_0.32.0        

loaded via a namespace (and not attached):
[1] lattice_0.20-38        bitops_1.0-6           grid_3.6.2             zlibbioc_1.32.0       
[5] Matrix_1.2-18          tools_3.6.2            RCurl_1.95-4.13        compiler_3.6.2        
[9] GenomeInfoDbData_1.2.2

temp1 <- readGAlignments("Desktop/test/again/Nnk_chip_rep3.ATfilt.combined.dm6.chr4.bam")

temp2 <- readGAlignmentPairs("Desktop/test/again/Nnk_chip_rep3.ATfilt.combined.dm6.chr4.bam")
# worked right after I copied the file into this location. 
# BUT after R crashes and I restart it and try it again, it does NOT work
# the the issue is sporadic - sometimes it works, sometimes it crashes R

here's the crazy long R-crash message my mac shows (it's a lot more verbose than the linux crash was....)

Process:               R [5060]
Path:                  /Applications/R.app/Contents/MacOS/R
Identifier:            org.R-project.R
Version:               R 3.6.2 GUI 1.70 El Capitan build (7735)
Code Type:             X86-64 (Native)
Parent Process:        ??? [1]
Responsible:           R [5060]
User ID:               504

Date/Time:             2020-01-29 17:54:41.837 -0800
OS Version:            Mac OS X 10.14.6 (18G2022)
Report Version:        12
Anonymous UUID:        C98D92BF-E276-1288-F466-4F0DA6071B06


Time Awake Since Boot: 21000 seconds

System Integrity Protection: enabled

Crashed Thread:        0  Dispatch queue: com.apple.main-thread

Exception Type:        EXC_ARITHMETIC (SIGFPE)
Exception Codes:       EXC_I386_DIV (divide by zero)
Exception Note:        EXC_CORPSE_NOTIFY

Termination Signal:    Floating point exception: 8
Termination Reason:    Namespace SIGNAL, Code 0x8
Terminating Process:   exc handler [5060]

Thread 0 Crashed:: Dispatch queue: com.apple.main-thread
0   Rsamtools.so                  	0x000000010c6c5ea3 Template::is_mate(bam1_t const*, bam1_t const*, unsigned int const*) const + 51
1   Rsamtools.so                  	0x000000010c6c5b59 Template::mate(std::__1::queue<std::__1::list<bam1_t const*, std::__1::allocator<bam1_t const*> >, std::__1::deque<std::__1::list<bam1_t const*, std::__1::allocator<bam1_t const*> >, std::__1::allocator<std::__1::list<bam1_t const*, std::__1::allocator<bam1_t const*> > > > >&, unsigned int const*) + 441
2   Rsamtools.so                  	0x000000010c6c55e8 BamIterator::mate_touched_templates() + 168
3   Rsamtools.so                  	0x000000010c6c71b4 BamFileIterator::iterate_inprogress(BGZF*) + 164
4   Rsamtools.so                  	0x000000010c6c1f6a BamIterator::yield(BGZF*, bam_mates_t*) + 90
5   Rsamtools.so                  	0x000000010c6c2763 samread_mate + 147
6   Rsamtools.so                  	0x000000010c6d949b _do_scan_bam + 619
7   Rsamtools.so                  	0x000000010c6d9a18 _scan_bam + 360
8   libR.dylib                    	0x00000001042a7f54 R_doDotCall + 2996
9   libR.dylib                    	0x00000001042a95f3 do_dotcall + 323 (dotcode.c:1252)
10  libR.dylib                    	0x00000001042dfc61 bcEval + 27617 (eval.c:6721)
11  libR.dylib                    	0x00000001042d894d Rf_eval + 445
12  libR.dylib                    	0x00000001042f58c3 forcePromise + 179 (eval.c:516)
13  libR.dylib                    	0x000000010430014c getvar + 828
14  libR.dylib                    	0x00000001042dcef4 bcEval + 15988 (eval.c:6517)
15  libR.dylib                    	0x00000001042d894d Rf_eval + 445
16  libR.dylib                    	0x00000001042f58c3 forcePromise + 179 (eval.c:516)
17  libR.dylib                    	0x000000010430014c getvar + 828
18  libR.dylib                    	0x00000001042dcef4 bcEval + 15988 (eval.c:6517)
19  libR.dylib                    	0x00000001042d894d Rf_eval + 445
20  libR.dylib                    	0x00000001042f58c3 forcePromise + 179 (eval.c:516)
21  libR.dylib                    	0x000000010430014c getvar + 828
22  libR.dylib                    	0x00000001042dcef4 bcEval + 15988 (eval.c:6517)
23  libR.dylib                    	0x00000001042d894d Rf_eval + 445
24  libR.dylib                    	0x00000001042f58c3 forcePromise + 179 (eval.c:516)
25  libR.dylib                    	0x000000010430014c getvar + 828
26  libR.dylib                    	0x00000001042dcef4 bcEval + 15988 (eval.c:6517)
27  libR.dylib                    	0x00000001042d894d Rf_eval + 445
28  libR.dylib                    	0x00000001042f7209 R_execClosure + 2153
29  libR.dylib                    	0x00000001042f610a Rf_applyClosure + 346 (eval.c:1716)
30  libR.dylib                    	0x00000001042dfd71 bcEval + 27889 (eval.c:6740)
31  libR.dylib                    	0x00000001042d894d Rf_eval + 445
32  libR.dylib                    	0x00000001042f7209 R_execClosure + 2153
33  libR.dylib                    	0x00000001042f610a Rf_applyClosure + 346 (eval.c:1716)
34  libR.dylib                    	0x00000001042dfd71 bcEval + 27889 (eval.c:6740)
35  libR.dylib                    	0x00000001042d894d Rf_eval + 445
36  libR.dylib                    	0x00000001042f7209 R_execClosure + 2153
37  libR.dylib                    	0x00000001042f610a Rf_applyClosure + 346 (eval.c:1716)
38  libR.dylib                    	0x00000001042dfd71 bcEval + 27889 (eval.c:6740)
39  libR.dylib                    	0x00000001042d894d Rf_eval + 445
40  libR.dylib                    	0x00000001042f7209 R_execClosure + 2153
41  libR.dylib                    	0x00000001042f610a Rf_applyClosure + 346 (eval.c:1716)
42  libR.dylib                    	0x00000001042dfd71 bcEval + 27889 (eval.c:6740)
43  libR.dylib                    	0x00000001042d894d Rf_eval + 445
44  libR.dylib                    	0x00000001042f7209 R_execClosure + 2153
45  libR.dylib                    	0x00000001042f610a Rf_applyClosure + 346 (eval.c:1716)
46  libR.dylib                    	0x00000001042dfd71 bcEval + 27889 (eval.c:6740)
47  libR.dylib                    	0x00000001042d894d Rf_eval + 445
48  libR.dylib                    	0x00000001042f7209 R_execClosure + 2153
49  libR.dylib                    	0x00000001042f7f43 R_execMethod + 611 (eval.c:1954)
50  methods.so                    	0x000000010fb698e7 R_dispatchGeneric + 2455
51  libR.dylib                    	0x000000010433dec3 do_standardGeneric + 675 (objects.c:1287)
52  libR.dylib                    	0x00000001042dfc61 bcEval + 27617 (eval.c:6721)
53  libR.dylib                    	0x00000001042d894d Rf_eval + 445
54  libR.dylib                    	0x00000001042f7209 R_execClosure + 2153
55  libR.dylib                    	0x00000001042f610a Rf_applyClosure + 346 (eval.c:1716)
56  libR.dylib                    	0x00000001042dfd71 bcEval + 27889 (eval.c:6740)
57  libR.dylib                    	0x00000001042d894d Rf_eval + 445
58  libR.dylib                    	0x00000001042f7209 R_execClosure + 2153
59  libR.dylib                    	0x00000001042f610a Rf_applyClosure + 346 (eval.c:1716)
60  libR.dylib                    	0x00000001042dfd71 bcEval + 27889 (eval.c:6740)
61  libR.dylib                    	0x00000001042d894d Rf_eval + 445
62  libR.dylib                    	0x00000001042f7209 R_execClosure + 2153
63  libR.dylib                    	0x00000001042f7f43 R_execMethod + 611 (eval.c:1954)
64  methods.so                    	0x000000010fb698e7 R_dispatchGeneric + 2455
65  libR.dylib                    	0x000000010433dec3 do_standardGeneric + 675 (objects.c:1287)
66  libR.dylib                    	0x00000001042dfc61 bcEval + 27617 (eval.c:6721)
67  libR.dylib                    	0x00000001042d894d Rf_eval + 445
68  libR.dylib                    	0x00000001042f7209 R_execClosure + 2153
69  libR.dylib                    	0x00000001042f610a Rf_applyClosure + 346 (eval.c:1716)
70  libR.dylib                    	0x00000001042dfd71 bcEval + 27889 (eval.c:6740)
71  libR.dylib                    	0x00000001042d894d Rf_eval + 445
72  libR.dylib                    	0x00000001042f7209 R_execClosure + 2153
73  libR.dylib                    	0x00000001042f7f43 R_execMethod + 611 (eval.c:1954)
74  methods.so                    	0x000000010fb698e7 R_dispatchGeneric + 2455
75  libR.dylib                    	0x000000010433dec3 do_standardGeneric + 675 (objects.c:1287)
76  libR.dylib                    	0x00000001042dfc61 bcEval + 27617 (eval.c:6721)
77  libR.dylib                    	0x00000001042d894d Rf_eval + 445
78  libR.dylib                    	0x00000001042f7209 R_execClosure + 2153
79  libR.dylib                    	0x00000001042f610a Rf_applyClosure + 346 (eval.c:1716)
80  libR.dylib                    	0x00000001042dfd71 bcEval + 27889 (eval.c:6740)
81  libR.dylib                    	0x00000001042d894d Rf_eval + 445
82  libR.dylib                    	0x00000001042f7209 R_execClosure + 2153
83  libR.dylib                    	0x00000001042f7f43 R_execMethod + 611 (eval.c:1954)
84  methods.so                    	0x000000010fb698e7 R_dispatchGeneric + 2455
85  libR.dylib                    	0x000000010433dec3 do_standardGeneric + 675 (objects.c:1287)
86  libR.dylib                    	0x00000001042dfc61 bcEval + 27617 (eval.c:6721)
87  libR.dylib                    	0x00000001042d894d Rf_eval + 445
88  libR.dylib                    	0x00000001042f7209 R_execClosure + 2153
89  libR.dylib                    	0x00000001042f610a Rf_applyClosure + 346 (eval.c:1716)
90  libR.dylib                    	0x00000001042d8e7d Rf_eval + 1773 (eval.c:743)
91  libR.dylib                    	0x00000001042fa41d do_set + 2125 (eval.c:2807)
92  libR.dylib                    	0x00000001042d8b40 Rf_eval + 944 (eval.c:695)
93  libR.dylib                    	0x0000000104329641 R_ReplDLLdo1 + 449 (main.c:393)
94  org.R-project.R               	0x0000000103e09ddf run_REngineRmainloop + 287
95  org.R-project.R               	0x0000000103dfde31 -[REngine runREPL] + 225
96  org.R-project.R               	0x0000000103decba1 main + 817
97  libdyld.dylib                 	0x00007fff594d23d5 start + 1

Thread 1:
0   libsystem_pthread.dylib       	0x00007fff596c53f0 start_wqthread + 0

Thread 2:: JavaScriptCore bmalloc scavenger
0   libsystem_kernel.dylib        	0x00007fff5960a866 __psynch_cvwait + 10
1   libsystem_pthread.dylib       	0x00007fff596c956e _pthread_cond_wait + 722
2   libc++.1.dylib                	0x00007fff56703a0a std::__1::condition_variable::wait(std::__1::unique_lock<std::__1::mutex>&) + 18
3   com.apple.JavaScriptCore      	0x00007fff30a2cc42 void std::__1::condition_variable_any::wait<std::__1::unique_lock<bmalloc::Mutex> >(std::__1::unique_lock<bmalloc::Mutex>&) + 82
4   com.apple.JavaScriptCore      	0x00007fff30a30d4b bmalloc::Scavenger::threadRunLoop() + 139
5   com.apple.JavaScriptCore      	0x00007fff30a30579 bmalloc::Scavenger::threadEntryPoint(bmalloc::Scavenger*) + 9
6   com.apple.JavaScriptCore      	0x00007fff30a31ee7 void* std::__1::__thread_proxy<std::__1::tuple<std::__1::unique_ptr<std::__1::__thread_struct, std::__1::default_delete<std::__1::__thread_struct> >, void (*)(bmalloc::Scavenger*), bmalloc::Scavenger*> >(void*) + 39
7   libsystem_pthread.dylib       	0x00007fff596c62eb _pthread_body + 126
8   libsystem_pthread.dylib       	0x00007fff596c9249 _pthread_start + 66
9   libsystem_pthread.dylib       	0x00007fff596c540d thread_start + 13

Thread 3:
0   libsystem_kernel.dylib        	0x00007fff5960e616 __select + 10
1   org.R-project.R               	0x0000000103def867 -[RController readThread:] + 647
2   com.apple.Foundation          	0x00007fff2f7a60e2 __NSThread__start__ + 1194
3   libsystem_pthread.dylib       	0x00007fff596c62eb _pthread_body + 126
4   libsystem_pthread.dylib       	0x00007fff596c9249 _pthread_start + 66
5   libsystem_pthread.dylib       	0x00007fff596c540d thread_start + 13

Thread 4:: com.apple.NSEventThread
0   libsystem_kernel.dylib        	0x00007fff5960721a mach_msg_trap + 10
1   libsystem_kernel.dylib        	0x00007fff59607768 mach_msg + 60
2   com.apple.CoreFoundation      	0x00007fff2d54b99e __CFRunLoopServiceMachPort + 328
3   com.apple.CoreFoundation      	0x00007fff2d54af0c __CFRunLoopRun + 1612
4   com.apple.CoreFoundation      	0x00007fff2d54a66e CFRunLoopRunSpecific + 455
5   com.apple.AppKit              	0x00007fff2ab484a2 _NSEventThread + 175
6   libsystem_pthread.dylib       	0x00007fff596c62eb _pthread_body + 126
7   libsystem_pthread.dylib       	0x00007fff596c9249 _pthread_start + 66
8   libsystem_pthread.dylib       	0x00007fff596c540d thread_start + 13

Thread 5:
0   libsystem_pthread.dylib       	0x00007fff596c53f0 start_wqthread + 0

Thread 6:
0   libsystem_pthread.dylib       	0x00007fff596c53f0 start_wqthread + 0

Thread 0 crashed with X86 Thread State (64-bit):
  rax: 0x0000000003541387  rbx: 0x0000000000000004  rcx: 0x00006000022ec580  rdx: 0x0000000000000000
  rdi: 0x0000600003724600  rsi: 0x000060000371b800  rbp: 0x00007ffeebe053c0  rsp: 0x00007ffeebe053a0
   r8: 0x000000000004fe5a   r9: 0x0000000003541387  r10: 0x00000000000034d0  r11: 0xffffffffffffffff
  r12: 0x0000000000000081  r13: 0x0000000000000001  r14: 0x000060000220f000  r15: 0x0000000000000851
  rip: 0x000000010c6c5ea3  rfl: 0x0000000000010246  cr2: 0x000000012fcd3000
  
Logical CPU:     0
Error Code:      0x00000000
Trap Number:     0


Binary Images:
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       0x103ef7000 -        0x103f21ff7 +libRblas.dylib (0) <CEB0747F-EC57-3430-A81F-E229A6C8B7BC> /Library/Frameworks/R.framework/Versions/3.6/Resources/lib/libRblas.dylib
       0x103f26000 -        0x104045fff +libgfortran.3.dylib (0) <53830B74-39EA-3EA7-8673-3CC9F041F6B7> /Library/Frameworks/R.framework/Versions/3.6/Resources/lib/libgfortran.3.dylib
       0x1040b1000 -        0x1040e8fe7 +libquadmath.0.dylib (0) <F6B458A7-8193-3831-B86E-DD832A5A19BE> /Library/Frameworks/R.framework/Versions/3.6/Resources/lib/libquadmath.0.dylib
       0x1040f7000 -        0x104121fd9 +libreadline.5.2.dylib (5.2) /Library/Frameworks/R.framework/Versions/3.6/Resources/lib/libreadline.5.2.dylib
       0x104135000 -        0x10414afef +libgcc_s.1.dylib (0) <20CA2D94-AB72-312A-ADD8-7912FE2574A0> /Library/Frameworks/R.framework/Versions/3.6/Resources/lib/libgcc_s.1.dylib
       0x104159000 -        0x1041c370f  dyld (655.1.1) <B3D9DDCB-5AA7-3674-9608-16B5AA4AA494> /usr/lib/dyld
       0x104224000 -        0x1044eefff +libR.dylib (0) <7E5EB822-A174-35A9-B626-8AAEAE85C797> /Library/Frameworks/R.framework/Versions/3.6/Resources/lib/libR.dylib
       0x1046ae000 -        0x104716fff +libomp.dylib (0) <5F2A02FB-2D20-3D6C-BEEE-40CA5EE3618C> /Library/Frameworks/R.framework/Versions/3.6/Resources/lib/libomp.dylib
       0x10619e000 -        0x1061a6ff3 +utils.so (0) <F4773C49-D219-3C39-B2E8-91DE6A437805> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/utils/libs/utils.so
       0x1069de000 -        0x1069e1047  libobjc-trampolines.dylib (756.2) <5795A048-3940-3801-90CE-33D1B1AF81F4> /usr/lib/libobjc-trampolines.dylib
       0x107dd3000 -        0x107dd5ff3 +bitops.so (0) <97B61BB0-6CB3-3045-BAAD-117408467B9A> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/bitops/libs/bitops.so
       0x107dd8000 -        0x107ddaff3 +lattice.so (0) <AF1D8EBC-EFCF-3431-B915-A8AD02714353> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/lattice/libs/lattice.so
       0x10b03d000 -        0x10b041ffb +Biobase.so (0) <910F69B3-87FE-3955-80D2-BC2D481EB269> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/Biobase/libs/Biobase.so
       0x10b045000 -        0x10b049ff7 +DelayedArray.so (0) <F658D4FF-F3B5-3A4D-B369-77B2FE942FF3> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/DelayedArray/libs/DelayedArray.so
       0x10b2b7000 -        0x10b2c5fff +BiocParallel.so (0) <BF940EEC-4FF0-3422-B117-9404AA6DF653> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/BiocParallel/libs/BiocParallel.so
       0x10b47c000 -        0x10b485ffb +RCurl.so (0) <A34226CB-90EC-3BD9-85B3-150AC5163FE4> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/RCurl/libs/RCurl.so
       0x10b728000 -        0x10b761ff7 +graphics.so (0) <0AA83284-0B38-3F36-AFA5-5D52AA5F7787> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/graphics/libs/graphics.so
       0x10b811000 -        0x10b81bffb +lapack.so (0) <FD42543E-FAED-384E-9238-143E5D5C3919> /Library/Frameworks/R.framework/Resources/modules/lapack.so
       0x10b822000 -        0x10b856ff3 +grDevices.so (0) <D7F27905-D7DD-3BA0-989F-E30012633D9C> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/grDevices/libs/grDevices.so
       0x10b8c1000 -        0x10b96bfff +stats.so (0) <0B71BE67-63E5-3F29-B3AD-8B151500979C> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/stats/libs/stats.so
       0x10c38b000 -        0x10c39cffb +tools.so (0) <4E4A55A5-B974-3FFA-961C-F1AB71811219> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/tools/libs/tools.so
       0x10c6b3000 -        0x10c763fff +Rsamtools.so (0) <8F52EFE1-9EB1-3D3A-A0CA-A83B73E70EA2> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/Rsamtools/libs/Rsamtools.so
       0x10caeb000 -        0x10caf4fff +GenomicAlignments.so (0) <C038A525-7CAB-30F4-9FD2-6155CEFFD097> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/GenomicAlignments/libs/GenomicAlignments.so
       0x10fb67000 -        0x10fb6cfff +methods.so (0) <01339C07-2C9C-38B2-B3A2-FDD55B73CC56> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/methods/libs/methods.so
       0x10fd25000 -        0x10fd29ff3 +parallel.so (0) <CA7EE20B-9E6A-3667-9B5F-D6EE12077089> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/parallel/libs/parallel.so
       0x10fd69000 -        0x10fd7cfff +IRanges.so (0) <26D70259-7288-3C38-B7BB-4B9DA415B200> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/IRanges/libs/IRanges.so
       0x10ff1e000 -        0x11011ffff +libRlapack.dylib (0) <61C208F7-2A76-35AC-97F1-B3F0CB57057A> /Library/Frameworks/R.framework/Versions/3.6/Resources/lib/libRlapack.dylib
       0x110193000 -        0x1101a4fff +XVector.so (0) <FA6186D7-E6FA-39F3-91CA-F06DBDA249F7> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/XVector/libs/XVector.so
       0x11045b000 -        0x110460ff7 +GenomicRanges.so (0) <7BBF99F9-59DA-37B8-B3FC-B83F5CE534D2> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/GenomicRanges/libs/GenomicRanges.so
       0x1105e2000 -        0x1105fcffb +grid.so (0) <E4EB1B95-5E8B-353F-B543-E8F00F5CE425> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/grid/libs/grid.so
       0x112a24000 -        0x112a40ff7 +S4Vectors.so (0) <D0A6522A-F114-36C0-A491-50B95D98385D> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/S4Vectors/libs/S4Vectors.so
       0x1132cc000 -        0x1134e4fff +matrixStats.so (0) <407BC54B-A660-3E8A-A4BD-1729EF91C9A5> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/matrixStats/libs/matrixStats.so
       0x113864000 -        0x113925ffb +Matrix.so (0) <2E7B3643-1BE7-3746-806A-F2738620947A> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/Matrix/libs/Matrix.so
       0x11d689000 -        0x11d6aeff7 +Biostrings.so (0) <C18502AB-FE8C-3544-9A3E-5DDFD8FC0AEC> /Library/Frameworks/R.framework/Versions/3.6/Resources/library/Biostrings/libs/Biostrings.so
    0x7fff21c73000 -     0x7fff21fccfff  com.apple.RawCamera.bundle (8.15.0 - 1031.4.4) <AB6E8A8F-0BFE-37EE-A135-44ABA4FCB559> /System/Library/CoreServices/RawCamera.bundle/Contents/MacOS/RawCamera
    0x7fff21fcf000 -     0x7fff220dbfff  com.apple.AMDMTLBronzeDriver (2.11.21 - 2.1.1) <D0034ABB-2AD7-3227-B34C-5038871E4D98> /System/Library/Extensions/AMDMTLBronzeDriver.bundle/Contents/MacOS/AMDMTLBronzeDriver
    0x7fff29548000 -     0x7fff29724ffb  com.apple.avfoundation (2.0 - 1550.4) <5854207B-6106-3DA4-80B6-36C42D042F26> /System/Library/Frameworks/AVFoundation.framework/Versions/A/AVFoundation
    0x7fff29725000 -     0x7fff297eafff  com.apple.audio.AVFAudio (1.0 - ???) <D454A339-2FC6-3EF6-992F-D676046612DB> /System/Library/Frameworks/AVFoundation.framework/Versions/A/Frameworks/AVFAudio.framework/Versions/A/AVFAudio
    0x7fff298f2000 -     0x7fff298f2fff  com.apple.Accelerate (1.11 - Accelerate 1.11) <762942CB-CFC9-3A0C-9645-A56523A06426> /System/Library/Frameworks/Accelerate.framework/Versions/A/Accelerate
    0x7fff298f3000 -     0x7fff29909ff7  libCGInterfaces.dylib (506.22) <1B6C92D9-F4B8-37BA-9635-94C4A56098CE> /System/Library/Frameworks/Accelerate.framework/Versions/A/Frameworks/vImage.framework/Versions/A/Libraries/libCGInterfaces.dylib
    0x7fff2990a000 -     0x7fff29fa3fef  com.apple.vImage (8.1 - ???) <53FA3611-894E-3158-A654-FBD2F70998FE> /System/Library/Frameworks/Accelerate.framework/Versions/A/Frameworks/vImage.framework/Versions/A/vImage
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  Calls made by this process:
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  Calls made by all processes on this machine:
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VM Region Summary:
ReadOnly portion of Libraries: Total=502.9M resident=0K(0%) swapped_out_or_unallocated=502.9M(100%)
Writable regions: Total=2.5G written=0K(0%) resident=0K(0%) swapped_out=0K(0%) unallocated=2.5G(100%)
 
                                VIRTUAL   REGION 
REGION TYPE                        SIZE    COUNT (non-coalesced) 
===========                     =======  ======= 
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CG backing stores                 35.5M        3 
CG image                           908K       19 
CoreAnimation                     92.0M       15 
CoreGraphics                         8K        1 
CoreImage                          376K       21 
CoreUI image data                 1660K       13 
CoreUI image file                  352K        4 
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JS JIT generated code              1.0G        3 
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MALLOC guard page                   32K        7 
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Memory Tag 242                      12K        1 
Memory Tag 251                      24K        2 
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Stack                             11.0M        7 
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WebKit Malloc                     8312K        5 
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__TEXT                           277.5M      382 
__UNICODE                          564K        1 
mapped file                       82.6M       46 
shared memory                     2680K        9 
===========                     =======  ======= 
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Model: iMac18,3, BootROM 179.0.0.0.0, 4 processors, Intel Core i5, 3.5 GHz, 16 GB, SMC 2.41f1
Graphics: kHW_AMDRadeonPro575Item, Radeon Pro 575, spdisplays_pcie_device, 4 GB
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USB Device: FaceTime HD Camera (Built-in)
USB Device: Keyboard Hub
USB Device: Apple Keyboard
Thunderbolt Bus: iMac, Apple Inc., 41.1

install error

Hi, I trid to install the package and met an error. My OS is Red Hat and GCC vresion is 8.3.0 and the package version is GenomicAlignments_1.10.1. I can't install the new version because of the limit of Rsamtools vresion could you help me fix this problem ? Thank you very much .

encodeOverlaps_methods.c: In function 'CharAE_append_int':
encodeOverlaps_methods.c:49:2: warning: implicit declaration of function 'append_string_to_CharAE' [-Wimplicit-function-declaration]
append_string_to_CharAE(char_ae, buf);
^~~~~~~~~~~~~~~~~~~~~~~
gcc -std=gnu99 -shared -fPIC -L/ds3512/home/panyp/ruanjian/packages/lib -o GenomicAlignments.so IRanges_stubs.o R_init_GenomicAlignments.o S4Vectors_stubs.o cigar_utils.o coordinate_mapping_methods.o encodeOverlaps_methods.o
installing to /ds3512/home/panyp/ruanjian/R-4.0.3/lib64/R/library/00LOCK-GenomicAlignments/00new/GenomicAlignments/libs
** R
** inst
** byte-compile and prepare package for lazy loading
Warning: undefined slot classes in definition of "GAlignments": NAMES(class "characterORNULL")
Error in rematchDefinition(definition, fdef, mnames, fnames, signature) :
methods can add arguments to the generic 'seqinfo<-' only if '...' is an argument to the generic
Error: unable to load R code in package 'GenomicAlignments'
Execution halted
ERROR: lazy loading failed for package 'GenomicAlignments'

  • removing '/ds3512/home/panyp/ruanjian/R-4.0.3/lib64/R/library/GenomicAlignments'

fail on r-devel

I have developed a package that uses GenomicAlignments and I am trying to submit my package to cran. However, the tests on r-devel-windows-ix86+x86_64 are failing with a backtrace that points to GenomicAlignments::readGAlignments. Can you please help?

My package:
https://github.com/alkodsi/ctDNAtools

The error:
test_check("ctDNAtools")
-- 1. Error: (unknown) (@test-ctDNAtest.R#13) ---------------------------------
trying to get slot "className" from an object of a basic class ("character") with no slots
Backtrace:
1. ctDNAtools::test_ctDNA(...)
2. ctDNAtools::get_mutations_read_names(...)
3. purrr::pmap(...)
4. ctDNAtools:::.f(...)
5. ctDNAtools:::get_mutation_read_names(...)
6. GenomicAlignments::stackStringsFromBam(...)
8. GenomicAlignments::readGAlignments(...)
10. GenomicAlignments::readGAlignments(...)
11. GenomicAlignments:::.bindExtraData(...)
15. methods::is(listData[[1L]], getClass("Annotated"))

Performance of readGAlignmentPairs with inter-chromosomal pairs

I'm contemplating switching over csaw to use GenomicAlignments for BAM file access, rather than continuing to wrestle with Rhtslib and its poor documentation/unreliability on Windows.

I've been looking at the readGAlignments and readGAlignmentPairs functions, which should be swap-in replacements for my main internal BAM reading functions (see LTLA/csaw#4). Currently, csaw runs on sorted BAM files on a chromosome-by-chromosome basis, so I would be calling these functions with which= set to an interval spanning the current chromosome in each iteration.

However, I am concerned about the performance implications of readGAlignmentPairs's obligations to retrieve mates from other chromosomes. Even with indexing, this would seem to require a lot of additional file accesses that would be unnecessary for my use case.

Would it be worthwhile to add an option to ignore paired reads that are not on the same reference sequence? Alternatively, this could feasibly be part of an expanded suite of scanBamParam options based on the INS isize field, e.g., keeping only non-zero INS isize values.

Implementation of queryLoc2refLoc() function

Is there a schedule for the implementation of those functions?

queryLocs2refLocs()
queryLoc2refLoc()

I saw a forum post from 4 years ago with the same question.
Do you take pull requests for this?
Best, Caspar

Clarify Counting of Query-Spanning Reads in Documentation

This example produces a count of 1 which I expected it to, but others might expect a count of zero, because the read spans the query coordinates and none of its matching bases fall within the query. I suppose that the query is typically much bigger (e.g. transcript) than the subject (e.g. read) unlike the example below (e.g. short variant).

GA <- GAlignments(seqnames=Rle("chr1"), pos=1L, cigar="50M10D50M", strand=Rle(factor('+', levels = c('+', '-', '*'))))
countOverlaps(GRanges("chr1", IRanges(51, 60)), GA)

Is there somewhere in the vignette that could provide a clarifying explanation in time for the next version?

When I use the readGAlignments .

image

When I used the software minimap2 to compare the bam files obtained by comparing ont data, I found a problem when I used this package to read the bam files. My main purpose was to obtain each read of the compared bam files. Start and stop site information in the reference genome.

'cigar_buf' overflow ERROR when calling GenomicAlignments::cigarNarrow()

test.cigar.txt

There is a 'cigar_buf' overflow ERROR when trying to subset a fairly large cigar string using GenomicAlignments::cigarNarrow function.

library(GenomicAlignments)

cg <- read.table(file = '<test.cigar.txt>')
cg <- as.character(cg)

Get matched bases

matches <- sum(GenomicAlignments::explodeCigarOpLengths(cigar = cg, ops = c('=','M'))[[1]])

Throws and 'cigar_buf' overflow ERROR

GenomicAlignments::cigarNarrow(cigar = cg, start = 1, end = matches)

Subsetting a shorter substring works

GenomicAlignments::cigarNarrow(cigar = cg, start = 1, end = 1000000)[[1]]

SessionInfo

sessionInfo()
R version 4.2.1 (2022-06-23)
Platform: x86_64-pc-linux-gnu (64-bit)
Running under: Ubuntu 20.04.3 LTS

Matrix products: default
BLAS: /usr/lib/x86_64-linux-gnu/blas/libblas.so.3.9.0
LAPACK: /usr/lib/x86_64-linux-gnu/lapack/liblapack.so.3.9.0

locale:
[1] LC_CTYPE=en_US.UTF-8 LC_NUMERIC=C LC_TIME=en_US.UTF-8 LC_COLLATE=en_US.UTF-8
[5] LC_MONETARY=en_US.UTF-8 LC_MESSAGES=en_US.UTF-8 LC_PAPER=en_US.UTF-8 LC_NAME=C
[9] LC_ADDRESS=C LC_TELEPHONE=C LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C

attached base packages:
[1] stats4 stats graphics grDevices utils datasets methods base

other attached packages:
[1] GenomicAlignments_1.34.0 Rsamtools_2.14.0 Biostrings_2.66.0 XVector_0.38.0
[5] SummarizedExperiment_1.28.0 Biobase_2.58.0 MatrixGenerics_1.10.0 matrixStats_0.63.0
[9] ggbio_1.46.0 ggplot2_3.4.0 GenomicRanges_1.50.2 GenomeInfoDb_1.34.4
[13] IRanges_2.32.0 S4Vectors_0.36.1 BiocGenerics_0.44.0 SVbyEye_0.0.0.9000

loaded via a namespace (and not attached):
[1] backports_1.4.1 Hmisc_4.7-2 BiocFileCache_2.6.0 plyr_1.8.8
[5] lazyeval_0.2.2 splines_4.2.1 BiocParallel_1.32.4 usethis_2.1.6
[9] digest_0.6.31 ensembldb_2.22.0 htmltools_0.5.4 wesanderson_0.3.6
[13] fansi_1.0.3 magrittr_2.0.3 checkmate_2.1.0 memoise_2.0.1
[17] BSgenome_1.66.1 cluster_2.1.4 remotes_2.4.2 ggfittext_0.9.1
[21] prettyunits_1.1.1 jpeg_0.1-10 colorspace_2.0-3 rappdirs_0.3.3
[25] blob_1.2.3 xfun_0.35 dplyr_1.0.10 callr_3.7.3
[29] crayon_1.5.2 RCurl_1.98-1.9 graph_1.76.0 VariantAnnotation_1.44.0
[33] survival_3.4-0 glue_1.6.2 polyclip_1.10-4 gtable_0.3.1
[37] zlibbioc_1.44.0 DelayedArray_0.24.0 pkgbuild_1.4.0 scales_1.2.1
[41] DBI_1.1.3 GGally_2.1.2 miniUI_0.1.1.1 Rcpp_1.0.9
[45] progress_1.2.2 xtable_1.8-4 htmlTable_2.4.1 foreign_0.8-84
[49] bit_4.0.5 OrganismDbi_1.40.0 Formula_1.2-4 profvis_0.3.7
[53] htmlwidgets_1.6.0 httr_1.4.4 RColorBrewer_1.1-3 ellipsis_0.3.2
[57] urlchecker_1.0.1 pkgconfig_2.0.3 reshape_0.8.9 XML_3.99-0.13
[61] farver_2.1.1 dbplyr_2.2.1 nnet_7.3-18 deldir_1.0-6
[65] utf8_1.2.2 tidyselect_1.2.0 labeling_0.4.2 rlang_1.0.6
[69] reshape2_1.4.4 later_1.3.0 AnnotationDbi_1.60.0 munsell_0.5.0
[73] tools_4.2.1 cachem_1.0.6 cli_3.4.1 generics_0.1.3
[77] RSQLite_2.2.19 devtools_2.4.5 evaluate_0.19 stringr_1.5.0
[81] fastmap_1.1.0 yaml_2.3.6 processx_3.8.0 knitr_1.41
[85] bit64_4.0.5 fs_1.5.2 purrr_0.3.5 KEGGREST_1.38.0
[89] AnnotationFilter_1.22.0 RBGL_1.74.0 mime_0.12 xml2_1.3.3
[93] biomaRt_2.54.0 compiler_4.2.1 rstudioapi_0.14 filelock_1.0.2
[97] curl_4.3.3 png_0.1-8 tibble_3.1.8 tweenr_2.0.2
[101] stringi_1.7.8 ps_1.7.2 GenomicFeatures_1.50.3 lattice_0.20-45
[105] ProtGenerics_1.30.0 Matrix_1.5-3 vctrs_0.5.1 pillar_1.8.1
[109] lifecycle_1.0.3 BiocManager_1.30.19 data.table_1.14.6 bitops_1.0-7
[113] httpuv_1.6.7 gggenes_0.4.1 rtracklayer_1.58.0 R6_2.5.1
[117] BiocIO_1.8.0 latticeExtra_0.6-30 promises_1.2.0.1 gridExtra_2.3
[121] sessioninfo_1.2.2 codetools_0.2-18 dichromat_2.0-0.1 MASS_7.3-58.1
[125] assertthat_0.2.1 pkgload_1.3.2 rjson_0.2.21 withr_2.5.0
[129] GenomeInfoDbData_1.2.9 hms_1.1.2 parallel_4.2.1 grid_4.2.1
[133] rpart_4.1.19 rmarkdown_2.19 biovizBase_1.46.0 ggnewscale_0.4.8
[137] ggforce_0.4.1 shiny_1.7.4 base64enc_0.1-3 interp_1.1-3
[141] restfulr_0.0.15

BiocParallel errors

Hi,
I'm getting this error when running readGAlignments on a BamViews object as follows:
readGAlignments(bamfiles,param=ScanBamParam(what=scanBamWhat(),tag=c("rq")),with.which_label=TRUE)

Error: BiocParallel errors
2 remote errors, element index: 1, 2
0 unevaluated and other errors
first remote error: cannot coerce class "list" to a DataFrame
In addition: Warning messages:
1: In readGAlignments(bamfiles, param = ScanBamParam(what = scanBamWhat(), :
'bamRanges(file)' and 'bamWhich(param)' differ; using 'bamRanges(file)'
2: stop worker failed:
wrong args for environment subassignment

summarizeOverlap performance issues and crashes.

I've been trying to debug some performance problems in DiffBind this week and I've traced them back to a few things in GenomicAlignemnts (called in diffbind through summarizeOverlaps). The issues I encountered with summarizeOverlaps are as follows:

  1. In some circumstances, providing a set of intervals twice, both to the 'features' argument and passed in through a 'ScanBamParam' object results in much faster performance (particularly using a small number of intervals e.g. just chromosome 1), which seems counterintuitive since the function should already 'know' which intervals to use.

  2. Using 'singleEnd=FALSE' seems to result in the entire file being read into memory, which is unexpected and can cause serious performance issues.

  3. Combining both of these and passing a set of intervals through ScanBamParam(which=intervals) and also specifying singleEnd=FALSE results in an unrecoverable hang, even in cases where the single-ended function runs in seconds.

Reproducible examples:

library(GenomicAlignments)
library(rtracklayer)

bamfile = "large_file.bam"

bed_file = "peaks_chr1.bed"

intervals = rtracklayer::import(bed_file)

# ISSUE 1
# this runs fine

t1 = system.time({
  counts = summarizeOverlaps(features=intervals,reads=bamfile,
                             ignore.strand=TRUE,singleEnd=TRUE)
})

# but this runs significantly faster, even though intervals are already provided
params = Rsamtools::ScanBamParam(which=intervals)

t2 = system.time({
  counts = summarizeOverlaps(features=intervals,reads=bamfile,
                             ignore.strand=TRUE,singleEnd=TRUE,
                             param=params)
})

# ISSUE 2
# this causes the entire file to be read into memory
counts = summarizeOverlaps(features=intervals,reads=bamfile,
                           ignore.strand=TRUE,singleEnd=FALSE)

# ISSUE 3
# this causes an unrecoverable hang, run at your own risk!
# counts = summarizeOverlaps(features=intervals,reads=bamfile,
#                            ignore.strand=TRUE,singleEnd=FALSE,
#                            param=params)

sessionInfo:

> sessionInfo()
R version 4.0.5 (2021-03-31)
Platform: x86_64-pc-linux-gnu (64-bit)
Running under: Ubuntu 20.04.2 LTS

Matrix products: default
BLAS:   /usr/lib/x86_64-linux-gnu/blas/libblas.so.3.9.0
LAPACK: /usr/lib/x86_64-linux-gnu/lapack/liblapack.so.3.9.0

locale:
 [1] LC_CTYPE=en_GB.UTF-8       LC_NUMERIC=C              
 [3] LC_TIME=en_GB.UTF-8        LC_COLLATE=en_GB.UTF-8    
 [5] LC_MONETARY=en_GB.UTF-8    LC_MESSAGES=en_GB.UTF-8   
 [7] LC_PAPER=en_GB.UTF-8       LC_NAME=C                 
 [9] LC_ADDRESS=C               LC_TELEPHONE=C            
[11] LC_MEASUREMENT=en_GB.UTF-8 LC_IDENTIFICATION=C       

attached base packages:
[1] parallel  stats4    stats     graphics  grDevices utils     datasets 
[8] methods   base     

other attached packages:
 [1] rtracklayer_1.50.0          DiffBind_3.1.7             
 [3] SummarizedExperiment_1.20.0 Biobase_2.50.0             
 [5] MatrixGenerics_1.2.1        matrixStats_0.58.0         
 [7] GenomicRanges_1.42.0        GenomeInfoDb_1.26.7        
 [9] IRanges_2.24.1              S4Vectors_0.28.1           
[11] BiocGenerics_0.36.1        

loaded via a namespace (and not attached):
  [1] backports_1.2.1          GOstats_2.56.0           BiocFileCache_1.14.0    
  [4] plyr_1.8.6               GSEABase_1.52.1          splines_4.0.5           
  [7] BiocParallel_1.24.1      ggplot2_3.3.3            amap_0.8-18             
 [10] digest_0.6.27            invgamma_1.1             GO.db_3.12.1            
 [13] SQUAREM_2021.1           fansi_0.4.2              magrittr_2.0.1          
 [16] checkmate_2.0.0          memoise_2.0.0            BSgenome_1.58.0         
 [19] base64url_1.4            limma_3.46.0             Biostrings_2.58.0       
 [22] annotate_1.68.0          systemPipeR_1.24.3       askpass_1.1             
 [25] bdsmatrix_1.3-4          prettyunits_1.1.1        jpeg_0.1-8.1            
 [28] colorspace_2.0-0         blob_1.2.1               rappdirs_0.3.3          
 [31] apeglm_1.12.0            ggrepel_0.9.1            dplyr_1.0.5             
 [34] crayon_1.4.1             RCurl_1.98-1.3           jsonlite_1.7.2          
 [37] graph_1.68.0             genefilter_1.72.1        brew_1.0-6              
 [40] survival_3.2-10          VariantAnnotation_1.36.0 glue_1.4.2              
 [43] gtable_0.3.0             zlibbioc_1.36.0          XVector_0.30.0          
 [46] DelayedArray_0.16.3      V8_3.4.1                 Rgraphviz_2.34.0        
 [49] scales_1.1.1             pheatmap_1.0.12          mvtnorm_1.1-1           
 [52] DBI_1.1.1                edgeR_3.32.1             Rcpp_1.0.6              
 [55] xtable_1.8-4             progress_1.2.2           emdbook_1.3.12          
 [58] bit_4.0.4                rsvg_2.1.1               AnnotationForge_1.32.0  
 [61] truncnorm_1.0-8          httr_1.4.2               gplots_3.1.1            
 [64] RColorBrewer_1.1-2       ellipsis_0.3.1           pkgconfig_2.0.3         
 [67] XML_3.99-0.6             dbplyr_2.1.1             locfit_1.5-9.4          
 [70] utf8_1.2.1               tidyselect_1.1.0         rlang_0.4.10            
 [73] AnnotationDbi_1.52.0     munsell_0.5.0            tools_4.0.5             
 [76] cachem_1.0.4             generics_0.1.0           RSQLite_2.2.7           
 [79] stringr_1.4.0            fastmap_1.1.0            yaml_2.2.1              
 [82] bit64_4.0.5              caTools_1.18.2           purrr_0.3.4             
 [85] RBGL_1.66.0              xml2_1.3.2               biomaRt_2.46.3          
 [88] compiler_4.0.5           rstudioapi_0.13          curl_4.3                
 [91] png_0.1-7                tibble_3.1.1             stringi_1.5.3           
 [94] GenomicFeatures_1.42.3   lattice_0.20-41          Matrix_1.3-2            
 [97] vctrs_0.3.7              pillar_1.6.0             lifecycle_1.0.0         
[100] irlba_2.3.3              data.table_1.14.0        bitops_1.0-7            
[103] R6_2.5.0                 latticeExtra_0.6-29      hwriter_1.3.2           
[106] ShortRead_1.48.0         KernSmooth_2.23-18       MASS_7.3-53.1           
[109] gtools_3.8.2             assertthat_0.2.1         openssl_1.4.3           
[112] Category_2.56.0          rjson_0.2.20             withr_2.4.2             
[115] GenomicAlignments_1.26.0 batchtools_0.9.15        Rsamtools_2.6.0         
[118] GenomeInfoDbData_1.2.4   hms_1.0.0                grid_4.0.5              
[121] DOT_0.1                  coda_0.19-4              GreyListChIP_1.22.0     
[124] ashr_2.2-47              mixsqp_0.3-43            bbmle_1.0.23.1          
[127] numDeriv_2016.8-1.1

I can provide the original files I used since they are public data but they are quite large, and I'd be interested to know if you can replicate them with different files. I've also attached my session info, which should be pretty up to date since this was a fresh install of both R and Bioconductor from 2 weeks ago.

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