PGS Documentation

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SubtitleTextIllumina Methylation BeadArray subworkflow
AnchorNameIllumina methylation workflow

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In the next dialog, use Browse… to point to the folder in which the .idat files were unzipped and use the Add File > button to move the .idat files to the box on the right (i.e. idat Files to Process; Figure 2). Select Next > to proceed.

 

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SubtitleTextUse the file browser to locate .idat files for import
AnchorNamefile browser

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The following dialog (Figure 3) deals with the manifest file, i.e. probe annotation. If a manifest file is not present locally, it will be downloaded in the Microarray libraries directory automatically. The download will take place in the background, with no particular message on the screen and it may take a few minutes, depending on the internet connection. If you have already downloaded the manifest file, to use it, select Use Available Manifest File option. If you have multiple versions available, you can pick an alternative by selecting Manually specify. The latter scenario corresponds to the situation where you want to reanalyze a data set at a later time. To enforce reproducibility, you may want to use the same version of the manifest file that you used when the data was first processed, rather than downloading an up-to-date version.

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SubtitleTextManifest File section of the import dialog lets you to specify the version of the manifest file (probe annotation) that you want to use. Output File specifies the location of the spreadsheet that will be created after the .idat files are imported
AnchorNameidat import dialog

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The Customize... button enables a granular control of the import process (Figure 4). First, the normalisation procedure is specified on the Algorithm tab (Figure 4).

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SubtitleTextSelecting the normalization transformation (functional normalization is the default)
AnchorNamenormalization options

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By default, import of .idat files will result in a single spreadsheet with beta values in the cells. The Outputs tab defines additional spreadsheets that can be created during data import (Figure 5).

 

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SubtitleTextAdvanced output options specifying additional spreadsheets that can be created during the .idat file import
AnchorNameadvnaced import output options

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Probe Intensity results in a spreadsheet with samples on rows (with suffix _probe added to the file name), probes on columns, and the sum of raw methylated and unmethylated probe intensities in the cells (the spreadsheet name will also have suffix _probe). Upon selecting Probe Intensity, you can additionally select Probe Signal, which will append rows to the "probe intensity" spreadsheet: for each sample a row with methylated (suffix _meth) and a separate row with unmethylated (suffix _unmeth) intensities (in the other words, three rows per sample)

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