"mogene10stv1probe" probe_length issue
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@debajyotibionova-14830
Last seen 6.9 years ago

Dear Concern, 

This is Deb from Hong Kong. I would like to bring your kind attention in connection with executing microarray data analysis with "mogene10stv1probe" (based on the original experimental facts). The range for this probe length is showing 21x25, which is supposed to be 25x25. Therefore, I am stuck with processing my data (.CEL files) using following command lines:

> probepkgname="mogene10stv1probe"  
> seq1=get(
probepkgname)
> prlen=nchar(seq1$sequence)
> range(
prlen)

[1] 21 25

> celfiles.gcrma <- gcrma(celfiles
Adjusting for optical effect...............................................................................................................................................................................................................................................................................................Done.
Computing affinitiesError: length(prlen) == 1 is not TRUE

## showing the above error message while computing affinities

However, I tried running one test run with human data, supported with "hgu133plus2". It was working perfectly. And,

> probepkgname="hgu133plus2 probe"  
> seq1=get(probepkgname)
> prlen=nchar(seq1$sequence)
> range(prlen)

[1] 25 25

## this range returned with 25x25 # thus, next commands for "adjusting optical effects" and "computing affinities" etc. are working smoothly. 

Please suggest me how to deal with this execution issue. Thanks in advance for your kind concern. 

Thank you. 

Best, 

Deb

bioconductor affymetrix mouse gene arrays mouse_gene st1.0 • 1.0k views
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@james-w-macdonald-5106
Last seen 6 hours ago
United States

You cannot in general use gcrma for the newer random primer arrays, as they are PM-only. You can point to some background probes to estimate the expected background level for each GC bin, but there probably aren't enough such probes to get a good estimate. In addition, you should be using the oligo or xps packages to analyze these arrays. Although the affy package has been modified to use these arrays, there are some probes that are shared between different probesets and the affy package cannot handle that situation correctly.

 

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Dear James, 

Thank you for your response. However, I used "oligo" packages specifically. But still no good returns. I used "digest", "affy", "simpleaffy", "genefilter". But no specific difference, I noticed. Can you please help me to specifically use some "affy" package that has been modified to accommodate these arrays as you mentioned above? If so, I will grateful. And, maybe further execution guideline! I am just stuck with this step and eagerly hunting for the solution.

Thank you. 

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Simply saying that you did something and didn't get 'good' results is not really meaningful, as what you think are 'good' results may not conform to what somebody else thinks that might mean, and it's not clear at all what you have done.

The basic paradigm is to do something like

library(oligo)

dat <- read.celfiles(list.celfiles())

eset <- rma(dat)

assuming of course that you have started R in the directory that contains the celfiles you wish to analyze. There is a vignette for the oligo package that you can consult, and for downstream analyses there is the limma User's Guide which is pretty comprehensive.

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Thanks James for your kind note. Yes, I agree your viewpoint of course. Anyway, I am gonna try this way that you have indicated. Thanks. 

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Entering edit mode
@debajyotibionova-14830
Last seen 6.9 years ago

Dear James, 

Thank you for your response. However, I used "oligo" packages specifically. But still no good returns. I used "digest", "affy", "simpleaffy", "genefilter". But no specific difference, I noticed. Can you please help me to specifically use some "affy" package that has been modified to accommodate these arrays as you mentioned above? If so, I will grateful. And, maybe further execution guideline! I am just stuck with this step and eagerly hunting for the solution.

Thank you. 

 

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