Error in reading data on isoformswitchanalyzeR
1
0
Entering edit mode
@saddamhusain77-19420
Last seen 5.7 years ago

I am trying to figure out how to use IsoformSwitchAnalyzeR on Salmon quantification. I'm really stuck at the very beginning. I have not been able to import the data. Below is the code I am using-

getwd()

[1] "/Users/saddamhusain/Desktop/salmon_isoswitch"

salmonQuant <- importIsoformExpression(

  • parentDir = system.file("salmonQuant", package="IsoformSwitchAnalyzeR")

  • )

Step 1 of 3: Identifying which algorithm was used...

Error in dirList[sapply(dirList, FUN = function(aDir) { :

invalid subscript type 'list'

salmon_isoswitch is a directory that contains salmonQuant subdirectory that in tern contains four independent subdirectories for each sample.

Any help will be appreciated.

isoformswitchanalyzeR rna-seq splice-variants • 2.7k views
ADD COMMENT
0
Entering edit mode

With this code-

salmonQuant <- importIsoformExpression("salmonQuant")

Step 1 of 3: Identifying which algorithm was used...

Error in importIsoformExpression("salmonQuant") : No directories of interest were found

Even though .txt files of count data are there.

ADD REPLY
0
Entering edit mode

Can you work through the example data successfully first?

https://bioconductor.org/packages/release/bioc/vignettes/IsoformSwitchAnalyzeR/inst/doc/IsoformSwitchAnalyzeR.html

I suggest reading the section in the longer workflow called ' Importing Data from Kallisto, Salmon, RSEM or StringTie'. It is pretty straightforward, I use this command, where this is simply the directory my salmon folders are in:

salmonQuant <- importIsoformExpression( parentDir = "C:/Users/chris/Desktop/isoswitcher/NewTranscriptomePEACbaseline/stringtie.salmon.v26/", addIsofomIdAsColumn = TRUE )

ADD REPLY
0
Entering edit mode

Thank you chris for the response. Yes, I can work through the example data smoothly but still getting the error with my data.

salmonQuant <- importIsoformExpression( parentDir = "/Users/saddamhusain/Desktop/salmon_isoswitch/")

Step 1 of 3: Identifying which algorithm was used...

Error in importIsoformExpression(parentDir = "/Users/saddamhusain/Desktop/salmon_isoswitch/") :

No directories of interest were found

salmonisoswitch folder contains four different folders named as "helc1" "helc2" "helt1" and "hel_t2" each of which contains respective quant.txt file.

ADD REPLY
0
Entering edit mode

OK, I think there is something different with your salmon output directories. I don't know what that is. Make sure your output directories contain all the files and folders normally outputted by salmon into a directory per sample.

ADD REPLY
0
Entering edit mode

If you are sure your output directories are as normal (they match example ones), then ask here for more help, https://github.com/kvittingseerup/IsoformSwitchAnalyzeR/issues.

ADD REPLY
0
Entering edit mode

I got quantification and gene quantification output from salmon ( v-0.13.1). I have done differential expression analysis on quantification files so I suppose same files should work out on isoformswitchanalyzer. Thanks for the suggestion. I have raised the same issue on GitHub too and am following it.

ADD REPLY
0
Entering edit mode

Try doing some example read alignments and then just trying to input the directory into R. I am having no problems with it, I'm using the latest version of salmon and not changing the output folder files and structure at all afterwards.

ADD REPLY
0
Entering edit mode

Thanks Chris, yeah, I will try to figure it out.

ADD REPLY
0
Entering edit mode
eshelden ▴ 40
@eshelden-14132
Last seen 5.6 years ago

The function importIsoformExpression in my version of the package has the following line: supportedTypes <- data.frame(orign = c("Kallisto", "Salmon", "RSEM", "StringTie"), fileName = c("abundance.tsv", "quant.sf", "isoforms.results", "tdata.ctab"), eLengthCol = c("efflength", "EffectiveLength", "effective_length", "")

I believe your Salmon file names need to end in ".quant.sf", not ".txt" in order to be considered valid.

ADD COMMENT

Login before adding your answer.

Traffic: 431 users visited in the last hour
Help About
FAQ
Access RSS
API
Stats

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.

Powered by the version 2.3.6