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openPrimeR #456

Closed matdoering closed 7 years ago

matdoering commented 7 years ago

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bioc-issue-bot commented 7 years ago

Hi @matdoering

Thanks for submitting your package. We are taking a quick look at it and you will hear back from us soon.

The DESCRIPTION file for this package is:

Package: openPrimeR
Title: Multiplex PCR Primer Design and Analysis
Version: 0.99.0
Authors@R: c(person("Matthias", "Döring", 
          email = "mdoering@mpi-inf.mpg.de", 
          role = c("aut", "cre")),
  person("Nico", "Pfeifer", 
          email = "pfeifer@informatik.uni-tuebingen.de", 
          role = c("aut"))
  )
Description: An implementation of methods for designing, evaluating,
    and comparing primer sets for multiplex PCR.
    Primers are designed by solving a set cover problem such that
    the number of covered template sequences is maximized with
    the smallest possible set of primers.
    To guarantee that high-quality primers are generated,
    only primers fulfilling constraints on their physicochemical properties 
    are selected. All relevant functions are accessible through
    a user-friendly Shiny application.
Depends:
    R (>= 3.4.0)
License: GPL-2
Encoding: UTF-8
LazyData: true
RoxygenNote: 6.0.1
Imports:
    Biostrings (>= 2.38.4),
    XML (>= 3.98-1.4),
    scales (>= 0.4.0),
    reshape2 (>= 1.4.1),
    seqinr (>= 3.3-3),
    IRanges (>= 2.4.8),
    GenomicRanges (>= 1.22.4),
    ggplot2 (>= 2.1.0),
    plyr (>= 1.8.4),
    dplyr (>= 0.5.0),
    stringdist (>= 0.9.4.1),
    stringr (>= 1.0.0),
    RColorBrewer (>= 1.1-2),
    DECIPHER (>= 1.16.1),
    lpSolveAPI (>= 5.5.2.0-17),
    digest (>= 0.6.9),
    Hmisc (>= 3.17-4),
    ape (>= 3.5),
    BiocGenerics (>= 0.16.1),
    S4Vectors (>= 0.8.11),
    foreach (>= 1.4.3),
    magrittr (>= 1.5),
    distr (>= 2.6),
    distrEx (>= 2.6),
    fitdistrplus (>= 1.0-7),
    uniqtag (>= 1.0),
    openxlsx (>= 4.0.17),
    grid (>= 3.1.0),
    grDevices (>= 3.1.0),
    stats (>= 3.1.0),
    utils (>= 3.1.0),
    methods (>= 3.1.0)
Suggests:
    shiny (>= 1.0.2),
    shinyjs (>= 0.9),
    shinyBS (>= 0.61),
    DT (>= 0.2),
    testthat (>= 1.0.2),
    knitr (>= 1.13),
    rmarkdown (>= 1.0),
    devtools (>= 1.12.0),
    doParallel (>= 1.0.10),
    pander (>= 0.6.0),
    learnr (>= 0.9)
SystemRequirements: MAFFT (>= 7.305),
    OligoArrayAux (>= 3.8),
    ViennaRNA (>= 2.2.4),
    MELTING (>= 5.1.1),
    Pandoc (>= 1.12.3)
biocViews: Software, Technology
VignetteBuilder: knitr
Collate: 
    'Comparison.R'
    'templates.R'
    'primers.R'
    'IO.R'
    'IO_view.R'
    'Ippolito.R'
    'RefCoverage.R'
    'Tiller.R'
    'ambiguity.R'
    'check_stop_codons.R'
    'con_annealing_temperature.R'
    'con_dimerization.R'
    'con_gc_clamp.R'
    'con_gc_ratio.R'
    'con_melting_temperature.R'
    'con_primer_coverage.R'
    'con_primer_efficiency.R'
    'con_primer_secondary_structures.R'
    'con_repeats.R'
    'con_runs.R'
    'con_template_secondary_structures.R'
    'constraints.R'
    'constraints_eval.R'
    'errors.R'
    'filters.R'
    'helper_functions.R'
    'initialize_primers.R'
    'initialize_primers_tree.R'
    'openPrimeR.R'
    'optimization_ILP.R'
    'optimization_algo.R'
    'optimization_global.R'
    'optimization_greedy.R'
    'plots_comparison.R'
    'settings.R'
    'plots_constraints.R'
    'plots_coverage.R'
    'plots_filtering.R'
    'primer_significance.R'
    'startApp.R'
    'zzz.R'
bioc-issue-bot commented 7 years ago

Your package has been approved for building. Your package is now submitted to our queue.

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bioc-issue-bot commented 7 years ago

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lawremi commented 7 years ago

Looks like there are a bunch of packages listed in Imports but not mentioned in NAMESPACE. Probably should just import(methods) instead of worrying about individual symbols. Lots of system dependencies. That's going to make the package tough to get working. Should at least consider separating the Shiny app into its own package.

bioc-issue-bot commented 7 years ago

Received a valid push; starting a build. Commits are:

25cef97 import of 'methods'

bioc-issue-bot commented 7 years ago

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On one or more platforms, the build results were: "skipped, ERROR". This may mean there is a problem with the package that you need to fix. Or it may mean that there is a problem with the build system itself.

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matdoering commented 7 years ago

Thanks for having a look! Your suggestion of making the Shiny app an individual package makes a lot of sense. Are there guidelines on how such "sister packages" should be provided? In my case I think it would be enough to ensure that the frontend package requires the correct version of the backend library but I'm wondering if there are other things to consider when doing a split.

Right now, I've just fixed the minor issues (data_raw folder, inst/doc removed) and imported 'methods'. Now, I'm wondering how to debug the OS-specific issues: if I do a version bump for every debug attempt that could be quite a lot of bumps to find out what the problem is. Is there some recommended way to debug OS-specificially if I don't have the OS available (e.g. OSX)? It's interesting that even for Linux the build doesn't work although it runs fine on my system (running R 3.4.0 and not 3.4.1, however).

bioc-issue-bot commented 7 years ago

Received a valid push; starting a build. Commits are:

028766f version bump

bioc-issue-bot commented 7 years ago

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On one or more platforms, the build results were: "skipped, ERROR". This may mean there is a problem with the package that you need to fix. Or it may mean that there is a problem with the build system itself.

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bioc-issue-bot commented 7 years ago

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bioc-issue-bot commented 7 years ago

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bioc-issue-bot commented 7 years ago

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bioc-issue-bot commented 7 years ago

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matdoering commented 7 years ago

I've just borrowed a Mac to identify the error that occurs during vignette building. In fact the error is thrown because GenomeInfoDbData does not seem to be available when doing a standard installation on a Mac using devtools::install(). After manually installing the package using biocLite() the vignette building worked fine. The problem is related to lazy evaluation since the vignette builder crashes the first time it encounters a call to GRanges, which requires GenomeInfoDb, which in turn requires GenomeInfoDbData. I've tried fixing the problem by including GenomeInfoDbData in the package DESCRIPTION but this also didn't help. The same thing happens on Travis-CI where the problem can be fixed by customizing the build procedure via manually requiring GenomeInfoDbData. A difference, however is, that Travis-CI throws the error sooner when checking whether the package dependencies are fulfilled following the installation and realizing that GenomeInfoDbData is not available.

To sum up, there seems to be a problem with correctly determining/installing the GenomeInfoDbData dependency for GenomicRanges on OSX. Any pointers?

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Your package has been built on Linux, Mac, and Windows.

On one or more platforms, the build results were: "skipped, ERROR". This may mean there is a problem with the package that you need to fix. Or it may mean that there is a problem with the build system itself.

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vobencha commented 7 years ago

Hi,

Your package should not have to depend on, include or suggest the GenomeInfoDbData data package. openPrimeR would need to depend / import the GenomeInfoDb software package only if you directly call functions from GenomeInfoDb.

openPrimeR imports GenomicRanges in DESCRIPTION but does not fully or selectively import it in the NAMESAPCE. That needs to be fixed. If you're using functions and classes from GenomicRanges only as internal helpers then importing is fine. If a user of openPrimeR needs access to functions from GenomicRanges to operate on objects defined in openPrimeR then GenomicRanges should go in depends in DESCRIPTION.

I don't know why we'd see a difference between mac and linux wrt lazy loading in this case. I haven't looked at your package code in detail - please try fixing the imports first and we'll go from there.

Valerie

matdoering commented 7 years ago

Dear Valerie, you're right about the GenomeInfoDb issue. The import in the NAMESPACE was just my failed attempt of trying to fix the OSX build error. Since then I've reverted back to the old DESCRIPTION with which Windows and Linux pass all checks, while OSX still fails. Is that OK for now in terms of passing the checks?

Could you explain the first part of your second paragraph to me?

openPrimeR imports GenomicRanges in DESCRIPTION but does not fully or selectively import it in the NAMESAPCE. That needs to be fixed.

I only use GenomicRanges with the double-colon operator (i.e. GenomicRanges::), which is why I never added any of its exported symbols to the NAMESPACE. And no, I don't use functions from GenomicRanges to access any objects defined in the package.

When I tested building the pkg with MacOS the error occurred here, the first time when calling the GRanges constructor in the foreach loop, complaining about GenomeInfoDbData not being installed.

vobencha commented 7 years ago

Historically we encouraged the use of selective imports in the NAMESPACE instead of the double colon operator, e.g.,

importFrom(GenomicRanges, GRanges)

Primary reasons were the accepted style at the time and clarity for what's being imported. With selective imports you can look at the NAMESPACE and quickly see what symbols the package was using. The current R documentation (https://cran.r-project.org/doc/manuals/r-release/R-exts.html#Specifying-imports-and-exports) seems to equate the two approaches. The 'imports' field in DESCRIPTION makes sure the package is available during load time so in theory all exported symbols will be available.

I won't push on this front - if you prefer to use the double colon operator that's fine. However I'd encourage you to be consistent. For example, Biostrings is in 'imports' in DESCRIPTION and one function (DNAStringSet) is selectively imported in the NAMESPACE. Then throughout the code you use '::' to call out many other functions from Biocstrings:

https://github.com/matdoering/openPrimeR/search?utf8=%E2%9C%93&q=Biostrings&type=

Either (1) fully import Biostrings in NAMESPACE or (2) selectively import from Biostrings in NAMESPACE or (3) don't import anything in NAMESPACE and use '::' for all functions from Biostrings.

I think the GenomeInfoDb problem may be related to R scoping rules in parallel evaluation. I would suggest printing out the environment inside the loop to see what packages/symbols are available. Related to parallel evaluation, it looks like in zzz.R you are circumventing parallel evaluation when on Windows. doParallel does support Windows ... was there a particular reason why you included this caveat?

I'll try testing on our Mac builder tomorrow. Valerie

matdoering commented 7 years ago

Thanks, I'll make sure that the imports are more consistent in the next version.

The GenomeInfoDb issue could be caused indeed be caused by the parallel call. Maybe the issue can already be solved by adding an importFrom(GenomicRanges, GRanges) to the NAMESPACE? I will test that in the next verison. I'd really appreciate it if you could look into it since I'm again without a Mac for testing.

The reason that parallel evaluations on Windows are disabled is that it it didn't work. Since it has been a couple of months when I encountered the issues, I cannot remember the exact error that was encountered but I can clearly recall that some evaluations just didn't work at all when the parallel backend was used. I can even remember that I tried different parallel implementations but encountered problems with all of them, which is why I disabled parallel computations on Windows by default.

I've worked on creating a separate package for the Shiny frontend, which can be found here. If I understand the Bioconductor documentation correctly, I can simply add this package here via AdditionalPackage once the initial package passes all checks, right?

Best Matthias

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matdoering commented 7 years ago

I've just uploaded a new version that doesn't contain the Shiny app. I tried fixing the MacOS problem by loading GRanges outside the foreach loop; this didn't help in fixing the problem though. Any help on this would be highly appreciated.

vobencha commented 7 years ago

For testing on the mac I first stangled the vignette


veracruz1:vignettes biocbuild$ R CMD Stangle openPrimeR_vignette.Rmd
Output file:  openPrimeR_vignette.R

Then sourced the vignette and got this error:


> source("openPrimeR
openPrimeR/            openPrimeR_vignette.R 
> source("openPrimeR_vignette.R")
There are missing/non-functioning external tools.
To use the full potential of openPrimeR, please make sure
that all of the listed tools are installed:
o MELTING (http://www.ebi.ac.uk/biomodels/tools/melting/)
o OligoArrayAux (http://unafold.rna.albany.edu/OligoArrayAux.php)
o MAFFT (http://mafft.cbrc.jp/alignment/software/)
The number of cores for was set to '2' by 'parallel_setup()'.

| Task             | Templates | Primers      | Input file format  |
|------------------|-----------|--------------|--------------------|
| Design primers   | ✓  |              | FASTA, CSV         |
| Analyze primers  | ✓  | ✓     | FASTA, CSV         |
| Compare primers  | ✓  | ✓     | FASTA, CSV         |
Reading settings file: C_Taq_PCR_high_stringency.xml
Due to missing external tools the following constraints were ignored:cross_dimerization,self_dimerization
Due to missing external tools the following coverage constraints were ignored:coverage_model

| Slot             | Getter/Setter |  Purpose |
|------------------|---------------|----------|
| `Input_Constraints` | `constraints()` | Desired values for primer properties |
| `Input_Constraint_Boundaries` | `constraintLimits()`  | Limits for relaxing constraints during primer design |
| `Coverage_Constraints` | `cvg_constraints()` | Constraints for estimating the coverage |
| `PCR_conditions` | `PCR()`  | Experimental PCR conditions |
| `constraint_settings` | `conOptions()` | Settings for evaluating the constraints |

# Phase 1: Initialization of primers
# Phase 2: Selection of primers
a) Filtering of primers
    o primer_length: 49 primers remaining (100%)
    o gc_ratio: 37 primers remaining (75.51%)
    o no_runs: 37 primers remaining (75.51%)
    o no_repeats: 37 primers remaining (75.51%)
    o melting_temp_range: 37 primers remaining (75.51%)
Error in out$Structure_fw[fw.idx] <- str.fw$Structure :
  replacement has length zero

The error is coming from this line in the vignette:


## ----design_primers, message = FALSE, warning = FALSE, eval = TRUE-------
# Design forward primers for the first two templates
optimal.primers <- design_primers(template.df[1:2,], mode.directionality = "fw",
                                  settings = design.settings)

Jump into the offending function with the debugger:


> debug(openPrimeR:::compute.secondary.structures)

and we see this error:


Error in read.secondary.structure.raw(out.file) :
  The input file: /var/folders/gs/3s3km1v97wg7qy7rnhs782680000gp/T//RtmpEG0y8F/viennaRNA_fw_d42e5188d82a04007b0924d4771abae3109da4d6c63d7.txt.fold does not exist.

The problem is that 'result' in this chunk of code is always NULL because 'out.file' does not exist:


    ...

    out.file <- paste0(out.file, ".fold")
        if (file.exists(out.file)) {
                result <- read.secondary.structure.raw(out.file)
        } else {
                result <- NULL
        }
    return(result)
}

Debugging in read.secondary.structure.raw I see that you expect RNAfold to append the word ".fold" to the output file. I am not familiar with ViennaRNA. Is it a guarantee that RNAfold always appends ".fold" ... and evidently it write this file directly to disk? It looks like you can pass an output file name to RNAfold which seems more robust than assuming the default will always remain the same. It looks like you've taken some precautions but please make sure you are (always) using tempfile() and tempdir() instead of writing directly to disk.

Valerie

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