PGS Documentation

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There are many useful visualizations, annotations, and biological interpretations interpretation tools that can operate on a gene list. In order for these features operate on work with an imported list, an annotation file must first be associated with the gene - list. Additionally, many operations that work with a list of significant genes (like GO- or Pathway-Enrichment) require comparison against a background of “non-significant” genes. The quickest way to accomplish both is to use the background of “all genes” for that organism provided by an annotation source like RefSeq, Ensembl, etc. in .pannot (Partek® annotationPartek annotation), .gff, .gtf, .bed, tab- or comma-delimited format. If the file is not already in a tab-separated or comma delimited format, you may import, modify, and save the file in the proper file format.

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SubtitleTextImporting an annotation file
AnchorNameImporting an annotation file

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  • Select the annotation file; we have selected in this example, we select a .pannot file downloaded from Partek distributed library file repository – hg19_refseq_14_01_03_v2.pannot from the C:/Microarry Libraries folder
  • Delete or rearrange the columns as necessary; we have placed the column with identifiers (should be unique ID) that correspond to our gene list first
  • Select (Image Removed) Select File then Save As Text File... to save the annotation file; we have named it Annotation File (Figure 2)

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SubtitleTextModified annotation file
AnchorNameModified Annotation File
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  • Select () to close the annotation file

Now we can add the annotation file to our imported gene list. 

  • Right click 1 (Gene Listgene_list.txt) in the spreadsheet tree
  • Select Properties from the pop-up menu

This brings up the Configure Genomic Properties dialog (Figure 3). 

 


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SubtitleTextSelecting an annotation file using the Configure Genomic Properties dialog
AnchorNameConfigure Genomic Properties

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  • Select Browse under Annotation File
  • Choose the annotation file; we have chosen Annotation File.txt txt 

If this is the first time you have used an annotation, the Configure Annotation dialog will launch. This is used to choose the columns with the chromosome number and position information for each feature. Our example annotation file has chromosome, start, and stop in separate columns. 

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  • Select Close to return to the Configure Genomic Properties dialogSelect appropriate
  • Select Set Column: to open the Choose column with gene symbols or microRNA names dialog (Figure 5)

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SubtitleTextChoosing the column in the annotation file with gene symbols or microRNA names
AnchorNameChoose column

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  • Select the appropriate column; here the default choice of 1. Symbol is appropriate
  • Select OK to return to the Configure Genomic Properties dialog
  • Select the appropriate species and genome build options; we have selected Homo sapiens and hg19
  • Select OK
  • hg19 (Figure 6)

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SubtitleTextThe gene list is now fully configured with an annotation file and reference genome selected
AnchorNameFully Configured Annotation

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  • Select OK
  • Select (Image Added) to save the spreadsheet 

The annotation file has been associated with the spreadsheet and additional tasks can now be performed on the data, e.g. since the annotation has genomic location, you can draw chromosome view on this data. 

Adding annotations to a spreadsheet 

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If an annotation file has been associated with a spreadsheet, annotations from the file can be added as columns in the spreadsheet when each identifier is on a row

  • Right click on a column header
  • Select Insert Annotation 
  • Select columns to add from Column Configuration  Configuration; we have selected Chromosome, Start, and Stop (Figure 57)
  • Select OK

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SubtitleTextAdding an annotation column from the annotation file
AnchorNameInserting Annotation

Annotating with cytobands

  • Select Annotate with Cytobands from Tools in the main toolbar when a suitable spreadsheet is open

A column with cytoband locations will be added to the spreadsheet. Adding a cytoband is possible if genomic coordinates are associated with the gene list spreadsheet during import or by association with an annotation file. 

Annotating with known SNPs

  • Select Annotate with Known SNPs from Tools in the main toolbar when a suitable spreadsheet is open will add 

A column of SNPs associated the listed genes and a column indicating the number of SNPs known to be associated with the genes will be added to the spreadsheet. If a SNP database has not been previously downloaded, it will need to be downloaded through the SNP database dialog (Figure 6). 

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SubtitleTextChoosing a database source for annotating a list of genes or genomic coordinates
AnchorNameAnnotating with a SNP database

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Alternatively, to generate a list of SNP IDs per row, right-click on a row header and select Create list of dbSNP.

In addition to SNPs, this feature can associate any data with a list of genes or genomic coordinates; the dbSNP database, any miRNA database, data from the Database of Genomic Variants (dgv), any mRNA transcriptome database, or any custom annotation source can be associated with your list. In each case, this feature will add columns to the imported gene list spreadsheet that match the genes with features from those databases.

 



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