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View Code? Open in Web Editor NEWMethylation and Expression AnaLysis
License: MIT License
Methylation and Expression AnaLysis
License: MIT License
Hi there,
I have tried MEAL for the analysis and integration of EPIC data and RNAseq but I have encountered an issue I am not able to sort it out.
I analyzed the methylation data using RnBeads
and I generated beta tables with beta values for all samples and probes non filtered out. Then, using the function minfi::makeGenomicRatioSetFromMatrix
and appropiate annotation (array = "IlluminaHumanMethylationEPIC
", annotation = "ilm10b4.hg19
") I create a GenomicRatio set.
> GRset_EPICs
class: GenomicRatioSet
dim: 705657 25
metadata(0):
assays(1): Beta
rownames(705657): cg14817997 cg26928153 ... cg07660283 cg09226288
rowData names(0):
colnames(25): AT102_VAT AT180_VAT ... CAT84_CF CAT86_CF
colData names(1): X1
Annotation
array: IlluminaHumanMethylationEPIC
annotation: ilm10b4.hg19
Preprocessing
Method: Matrix converted with makeGenomicRatioSetFromMatrix
minfi version: NA
Manifest version: NA
On the other hand, I analyzed the RNAseq using DESeq2 and from the dds object generated I retrieved normalized counts and using BioMart I retrieved genome coordinates.
> expr_set
ExpressionSet (storageMode: lockedEnvironment)
assayData: 16255 features, 28 samples
element names: exprs
protocolData: none
phenoData
sampleNames: AT102_VAT AT180_VAT ... CAT86_CF (28 total)
varLabels: Individual Gender ... agegroup (14 total)
varMetadata: labelDescription
featureData
featureNames: ENSG00000000003 ENSG00000000005 ... ENSG00000278845
(16255 total)
fvarLabels: chromosome start end strand
fvarMetadata: labelDescription
experimentData: use 'experimentData(object)'
Annotation:
I create a multidataset object:
multi <- createMultiDataSet()
multi <- add_genexp(multi, expr_set)
multi <- add_methy(multi, GRset_EPICs)
and check it out:
> commonSamples(multi)
Object of class 'MultiDataSet'
. assayData: 2 elements
. expression: 16255 features, 25 samples
. methylation: 705657 features, 25 samples
. featureData:
. expression: 16255 rows, 4 cols (chromosome, ..., end)
. methylation: 705657 rows, 5 cols (seqnames, ..., width)
. rowRanges:
. expression: YES
. methylation: YES
. phenoData:
. expression: 25 samples, 15 cols (Individual, ..., agegroup)
. methylation: 25 samples, 2 cols (X1, id)
When I tried to do the expression methylation correlation I failed and the error produced said:
> methExprs <- correlationMethExprs(multi)
Warning messages:
1: In .Seqinfo.mergexy(x, y) :
The 2 combined objects have no sequence levels in common. (Use
suppressWarnings() to suppress this warning.)
2: In correlationMethExprs(multi.filt) :
There are no expression probes in the range of the cpgs. An empty data.frame will be returned.
I wonder where the error is or what should I do to fix it. Might it be due to the annotation of the EPIC data? I wonder if you have any experience in this type of data. All examples I have seen from MEAL use 450K microarrays but I hope it also work with this new version of microarray data.
Thanks in advance,
Jose
Regards from IGTP
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