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START App

This is the code to run the app described in the manuscript:

Nelson, JW, Sklenar J, Barnes AP, Minnier J. (2016) "The START App: A Web-Based RNAseq Analysis and Visualization Resource." Bioinformatics. doi: 10.1093/bioinformatics/btw624.

The app is hosted on Shinyapps.io here: https://kcvi.shinyapps.io/START/

To run this app locally on your machine, download R or RStudio and run the following commands once to set up the environment:


install.packages(c("reshape2","ggplot2","ggthemes","gplots","ggvis","dplyr","tidyr","DT", "readr",
                   "RColorBrewer","pheatmap","shinyBS","plotly","janitor",
                   "markdown","NMF","scales","heatmaply"))
## try http:// if https:// URLs are not supported
if (!requireNamespace("BiocManager", quietly = TRUE))
  install.packages("BiocManager")
BiocManager::install(c("limma","edgeR"))

You may now run the shiny app with just one command in R:

shiny::runGitHub("STARTapp", "jminnier")

Jonathan Nelson, Jiri Sklenar, Anthony Barnes, Jessica Minnier. The Knight Cardiovascular Institute and OHSU-PSU School of Public Health, Oregon Health & Science University, Portland, OR 97239-3098, USA.

We would appreciate reports of any issues with the app via the issues option of Github or by emailing start.app.help-at-gmail.com.

Instructions

Instructions can be found here: https://github.com/jminnier/STARTapp/blob/master/instructions/Instructions.md

Licensing

This shiny code is licensed under the GPLv3. Please see the file LICENSE.txt for information.

START (Shiny Transcriptome Analysis Resource Tool) App
Shiny App for analysis and visualization of transcriptome data.
Copyright (C) 2016  Jessica Minnier

This program is free software: you can redistribute it and/or modify
it under the terms of the GNU General Public License as published by
the Free Software Foundation, either version 3 of the License, or
(at your option) any later version.

This program is distributed in the hope that it will be useful,
but WITHOUT ANY WARRANTY; without even the implied warranty of
MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE.  See the
GNU General Public License for more details.

You should have received a copy of the GNU General Public License
along with this program.  If not, see <http://www.gnu.org/licenses/>.

You may contact the author of this code, Jessica Minnier, at <[email protected]>

Code adapted for use in app:

Law, CW, Chen, Y, Shi, W, and Smyth, GK (2014). Voom: precision weights unlock linear model analysis tools for RNA-seq read counts. Genome Biology 15, R29.

Love MI, Huber W, and Anders S (2014). Moderated estimation of fold change and dispersion for RNA-Seq data with DESeq2. Genome Biology, 15, 550.

DOI

DOI

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startapp's Issues

Help regarding Shiny Transcriptome Analysis Resource Tool

Hello STARTapp team,

I am using the Shiny Transcriptome Analysis Resource webTool, available at

https://kcvi.shinyapps.io/START/#dataformats

It's the best tool that I have come across to analyze transcriptome data. Thanks for such a valuable webtool.

I need a small help with this app. I have made the PCA plot using this webtool. If I directly copy the PCA made on the website by right clicking and saving it as image the resolution is very low. The journals are not accepting this image, is there a better way to either derive or download a high resolution PCA plot using this tool.

My knowledge with R studio is very limited and I ma not being able to run this on there. SO my only option is using webtool.

I would really appriciate a quick response as I am have submitted my paper to the journal but they want a high resolution image asap.

Thanks in advance.

Regards

Shwetank, Ph.D.
Prof. Aron Lukacher's Lab,
Dept of Microbiology and Immunology,
Penn State College of Medicine
Hershey, PA 17033
Lab number : 717 531 6340

Input error

Hi

I am trying to run this app and am running into some trouble when i input the example data (exampleanalysisres_short.csv). My R Studio session just freezes. I am running it on a google chrome browser rather than the R-studio "browser", where it just kills it!

Since the file does not have p-vals, but rather adjp-vals, i am just using the same values from the files for both the inputs. Is this causing the freeze up?

Once i submit the files, it just greys over and i get a little "analyzing data .."
Any help would be appreciated

Thanks

Error below:
R version 3.5.1 (2018-07-02)
Platform: x86_64-w64-mingw32/x64 (64-bit)
Running under: Windows >= 8 x64 (build 9200)

Matrix products: default

locale:
[1] LC_COLLATE=English_United States.1252 LC_CTYPE=English_United States.1252
[3] LC_MONETARY=English_United States.1252 LC_NUMERIC=C
[5] LC_TIME=English_United States.1252

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

other attached packages:
[1] data.table_1.11.6 colourpicker_1.0 readr_1.1.1 heatmaply_0.15.2
[5] viridis_0.5.1 viridisLite_0.3.0 scales_1.0.0 NMF_0.21.0
[9] Biobase_2.40.0 BiocGenerics_0.26.0 cluster_2.0.7-1 rngtools_1.3.1
[13] pkgmaker_0.27 registry_0.5 markdown_0.8 plotly_4.8.0
[17] shinyBS_0.61 pheatmap_1.0.10 RColorBrewer_1.1-2 edgeR_3.22.4
[21] limma_3.36.5 DT_0.4 tidyr_0.8.1 dplyr_0.7.6
[25] ggvis_0.4.3 gplots_3.0.1 ggthemes_4.0.1 ggplot2_3.0.0
[29] reshape2_1.4.3 shiny_1.1.0

loaded via a namespace (and not attached):
[1] bitops_1.0-6 webshot_0.5.0 doParallel_1.0.14 httr_1.3.1
[5] prabclus_2.2-6 tools_3.5.1 R6_2.2.2 KernSmooth_2.23-15
[9] lazyeval_0.2.1 colorspace_1.3-2 trimcluster_0.1-2.1 nnet_7.3-12
[13] withr_2.1.2 tidyselect_0.2.4 gridExtra_2.3 compiler_3.5.1
[17] TSP_1.1-6 diptest_0.75-7 caTools_1.17.1.1 DEoptimR_1.0-8
[21] mvtnorm_1.0-8 robustbase_0.93-3 stringr_1.3.1 digest_0.6.17
[25] pkgconfig_2.0.2 htmltools_0.3.6 bibtex_0.4.2 htmlwidgets_1.2
[29] rlang_0.2.2 rstudioapi_0.7 BiocInstaller_1.30.0 bindr_0.1.1
[33] jsonlite_1.5 crosstalk_1.0.0 mclust_5.4.1 gtools_3.8.1
[37] dendextend_1.8.0 magrittr_1.5 modeltools_0.2-22 Rcpp_0.12.18
[41] munsell_0.5.0 stringi_1.2.4 whisker_0.3-2 yaml_2.2.0
[45] MASS_7.3-50 flexmix_2.3-14 plyr_1.8.4 grid_3.5.1
[49] gdata_2.18.0 promises_1.0.1 crayon_1.3.4 miniUI_0.1.1.1
[53] lattice_0.20-35 hms_0.4.2 locfit_1.5-9.1 pillar_1.3.0
[57] fpc_2.1-11.1 codetools_0.2-15 stats4_3.5.1 glue_1.3.0
[61] gclus_1.3.1 httpuv_1.4.5 foreach_1.4.4 gtable_0.2.0
[65] purrr_0.2.5 kernlab_0.9-27 assertthat_0.2.0 gridBase_0.4-7
[69] mime_0.5 xtable_1.8-3 later_0.7.5 class_7.3-14
[73] seriation_1.2-3 tibble_1.4.2 iterators_1.0.10 bindrcpp_0.2.2
[1] "inputting data"
Parsed with column specification:
cols(
Geneid = col_character(),
MGIsymbol = col_character(),
group1_1 = col_integer(),
group1_2 = col_integer(),
group1_3 = col_integer(),
group2_1 = col_integer(),
group2_2 = col_integer(),
group2_3 = col_integer(),
group3_1 = col_integer(),
group3_2 = col_integer(),
group3_3 = col_integer()
)
[1] "uploaded mousecounts data"
[1] "server-datafilter-update-filters"
[1] "analysisCountDataReactive"
[1] "server-datafilter-update-tests"
[1] "server-datafilter-update-expr"
[1] "server-dotplot-update"
[1] "server-heatmap-update"
[1] "server-samplegroupplots-update-yname"
[1] "server-samplegroupplots-update-groups"
[1] "server-sampleplots-update-samples"
[1] "server-analysisres-update"
[1] "drawing volcano plot"
[1] "drawing scatterplot"
[1] "inputting data"
Parsed with column specification:
cols(
Geneid = col_character(),
MGIsymbol = col_character(),
group1_1 = col_integer(),
group1_2 = col_integer(),
group1_3 = col_integer(),
group2_1 = col_integer(),
group2_2 = col_integer(),
group2_3 = col_integer(),
group3_1 = col_integer(),
group3_2 = col_integer(),
group3_3 = col_integer()
)
[1] "uploaded mousecounts data"
[1] "server-datafilter-update-filters"
[1] "analysisCountDataReactive"
[1] "server-datafilter-update-tests"
[1] "server-datafilter-update-expr"
[1] "server-dotplot-update"
[1] "server-heatmap-update"
[1] "server-samplegroupplots-update-yname"
[1] "server-samplegroupplots-update-groups"
[1] "server-sampleplots-update-samples"
[1] "server-analysisres-update"
[1] "drawing volcano plot"
[1] "drawing scatterplot"
[1] "inputting data"
[1] "server-datafilter-update-tests"
[1] "server-datafilter-update-expr"
[1] "server-sampleplots-update-samples"
[1] "server-datafilter-update-tests"
[1] "server-datafilter-update-expr"
[1] "server-sampleplots-update-samples"
[1] "updating analyzed data choices"
[1] "inputting data"
Parsed with column specification:
cols(
.default = col_double(),
Geneid = col_character(),
MGIsymbol = col_character()
)
See spec(...) for full column specifications.
[1] "uploaded seqdata"
[1] "server-analysisres-update"
[1] "analysisCountDataReactive"
Warning in if (input == "") { :
the condition has length > 1 and only the first element will be used
[1] "Num genes kept after removing empty geneids: 100 of 100"
Warning: Setting row names on a tibble is deprecated.
Warning: Setting row names on a tibble is deprecated.
Joining, by = c("unique_id", "test")
Joining, by = c("unique_id", "test")
Joining, by = "sampleid"
Warning: Column sampleid joining character vector and factor, coercing into character vector
[1] "drawing volcano plot"
[1] "drawing scatterplot"
[1] "server-datafilter-update-filters"
[1] "server-datafilter-update-tests"
Warning in max(tmpfc, na.rm = T) :
no non-missing arguments to max; returning -Inf
[1] "server-datafilter-update-expr"
Warning: Error in FUN: only defined on a data frame with all numeric variables
50: stop
49: FUN
48: lapply
47: Summary.data.frame
46: [server-filterdata.R#77]
3: runApp
2: runUrl
1: shiny::runGitHub
[1] "server-sampleplots-update-samples"
[1] "server-dotplot-update"
[1] "server-heatmap-update"
[1] "server-samplegroupplots-update-yname"
[1] "server-samplegroupplots-update-groups"

Filter by expression count button

Hello,
during the filterdata step, I can not filter the expression value by count, it goes directly to log2cpm_voom.
I can not select count.

Thanks,

Mohamed

Website won't load

Hello,

This is a great tool, and I use it in an assignment for a bioinformatics class I teach; however, this past week several student and myself are unable to load the website from https://kcvi.shinyapps.io/START/. It just hangs and says "Please Wait".

error after file submission

Hi,
when I press the submit button the app crashes and it complains:
[1] "server-sampleplots-update-samples"
Warning: Error in [: subscript out of bounds
94: extract_count_data [fun-input-analyze-data.R#30]
93: analyze_expression_data [fun-input-analyze-data.R#94]
89: eventReactiveHandler [server-inputdata.R#122]
45: analyzeDataReactive
44: [server-filterdata.R#56]
1: runApp

My input format is according to what has been described in the instructions. Is it because I have zero in some replicates.

Error: non-character argument, Volcano Plot

I tried following the installation instructions. The app works and I can upload csv data, but when I goto Analysis Plots, I get "Error: non-character argument". I even tried the example csv data, with the same result. Please advise.

This is the stack trace in the console:

Warning: Error in strsplit: non-character argument
Stack trace (innermost first):
87: strsplit
86: do.call
85: rna_volcanoplot [fun-analysisres.R#97]
84: eval [server-analysisres.R#76]
83: eval
82: withProgress
81: "plotly"::"ggplotly" [server-analysisres.R#67]
80: func
79: origRenderFunc
78: output$volcanoplot
3: runApp
2: runUrl
1: shiny::runGitHub

App crashes when a saved Rdata file is loaded

We noticed that the Rdata file produced from the current version is different from the previous version. However, when the saved Rdata file generated from the current version is uploaded to the same current version crashes the application.

non-integers causes crash with edgeR option

a comment by Les K said that this fixes and issue which makes me think it's the whole numbers (0s should be fine):

I tried a modification of the input data file and now the edgeR analysis works fine.
Namely, I made sure all the numbers were whole, no decimal places, and also added 1 to all numbers to get rid of zero values. I did both so I don’t know if one of the changes alone solved the problem.

adding statistics to the boxplots

Hello,
I have the raw read counts and I generated the boxplots with your application on NASQAR. The boxplots look beautiful, however, I don't see option to add statistics or p values for the purpose of statistical differentiation between two groups.

Could you please help me to find a way to add statistics?

Warning: Error in ggfun("create_layout"): unused argument (plot$coordinates)

hello,

I tried following the installation instructions. The app works and I can upload csv data, but when I goto Analysis Plots, I get Warning: Error in ggfun("create_layout"): unused argument (plot$coordinates). The example csv data, has the same result. Please advise.

Warning: Error in ggfun("create_layout"): unused argument (plot$coordinates)
Stack trace (innermost first):
93: gg2list
92: ggplotly.ggplot
91: ggplotly
90: plotly_build
89: plotly_build.gg
88: plotly_build
87: rna_scatterplot [fun-analysisres.R#196]
86: eval [server-analysisres.R#108]
85: eval
84: withProgress
83: "plotly"::"ggplotly" [server-analysisres.R#105]
82: func
81: origRenderFunc
80: output$scatterplot
3: runApp
2: runUrl
1: shiny::runGitHub

R version 3.4.4 (2018-03-15)
Platform: x86_64-pc-linux-gnu (64-bit)
Running under: Ubuntu 14.04.5 LTS

Many thanks

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