Partek
  • Overview
  • Partek Flow
    • Frequently Asked Questions
      • General
      • Visualization
      • Statistics
      • Biological Interpretation
      • How to cite Partek software
    • Quick Start Guide
    • Installation Guide
      • Minimum System Requirements
      • Single Cell Toolkit System Requirements
      • Single Node Installation
      • Single Node Amazon Web Services Deployment
      • Multi-Node Cluster Installation
      • Creating Restricted User Folders within the Partek Flow server
      • Updating Partek Flow
      • Uninstalling Partek Flow
      • Dependencies
      • Docker and Docker-compose
      • Java KeyStore and Certificates
      • Kubernetes
    • Live Training Event Recordings
      • Bulk RNA-Seq Analysis Training
      • Basic scRNA-Seq Analysis & Visualization Training
      • Advanced scRNA-Seq Data Analysis Training
      • Bulk RNA-Seq and ATAC-Seq Integration Training
      • Spatial Transcriptomics Data Analysis Training
      • scRNA and scATAC Data Integration Training
    • Tutorials
      • Creating and Analyzing a Project
        • Creating a New Project
        • The Metadata Tab
        • The Analyses Tab
        • The Log Tab
        • The Project Settings Tab
        • The Attachments Tab
        • Project Management
        • Importing a GEO / ENA project
      • Bulk RNA-Seq
        • Importing the tutorial data set
        • Adding sample attributes
        • Running pre-alignment QA/QC
        • Trimming bases and filtering reads
        • Aligning to a reference genome
        • Running post-alignment QA/QC
        • Quantifying to an annotation model
        • Filtering features
        • Normalizing counts
        • Exploring the data set with PCA
        • Performing differential expression analysis with DESeq2
        • Viewing DESeq2 results and creating a gene list
        • Viewing a dot plot for a gene
        • Visualizing gene expression in Chromosome view
        • Generating a hierarchical clustering heatmap
        • Performing biological interpretation
        • Saving and running a pipeline
      • Analyzing Single Cell RNA-Seq Data
      • Analyzing CITE-Seq Data
        • Importing Feature Barcoding Data
        • Data Processing
        • Dimensionality Reduction and Clustering
        • Classifying Cells
        • Differentially Expressed Proteins and Genes
      • 10x Genomics Visium Spatial Data Analysis
        • Start with pre-processed Space Ranger output files
        • Start with 10x Genomics Visium fastq files
        • Spatial data analysis steps
        • View tissue images
      • 10x Genomics Xenium Data Analysis
        • Import 10x Genomics Xenium Analyzer output
        • Process Xenium data
        • Perform Exploratory analysis
        • Make comparisons using Compute biomarkers and Biological interpretation
      • Single Cell RNA-Seq Analysis (Multiple Samples)
        • Getting started with the tutorial data set
        • Classify cells from multiple samples using t-SNE
        • Compare expression between cell types with multiple samples
      • Analyzing Single Cell ATAC-Seq data
      • Analyzing Illumina Infinium Methylation array data
      • NanoString CosMx Tutorial
        • Importing CosMx data
        • QA/QC, data processing, and dimension reduction
        • Cell typing
        • Classify subpopulations & differential expression analysis
    • User Manual
      • Interface
      • Importing Data
        • SFTP File Transfer Instructions
        • Import single cell data
        • Importing 10x Genomics Matrix Files
        • Importing and Demultiplexing Illumina BCL Files
        • Partek Flow Uploader for Ion Torrent
        • Importing 10x Genomics .bcl Files
        • Import a GEO / ENA project
      • Task Menu
        • Task actions
        • Data summary report
        • QA/QC
          • Pre-alignment QA/QC
          • ERCC Assessment
          • Post-alignment QA/QC
          • Coverage Report
          • Validate Variants
          • Feature distribution
          • Single-cell QA/QC
          • Cell barcode QA/QC
        • Pre-alignment tools
          • Trim bases
          • Trim adapters
          • Filter reads
          • Trim tags
        • Post-alignment tools
          • Filter alignments
          • Convert alignments to unaligned reads
          • Combine alignments
          • Deduplicate UMIs
          • Downscale alignments
        • Annotation/Metadata
          • Annotate cells
          • Annotation report
          • Publish cell attributes to project
          • Attribute report
          • Annotate Visium image
        • Pre-analysis tools
          • Generate group cell counts
          • Pool cells
          • Split matrix
          • Hashtag demultiplexing
          • Merge matrices
          • Descriptive statistics
          • Spot clean
        • Aligners
        • Quantification
          • Quantify to annotation model (Partek E/M)
          • Quantify to transcriptome (Cufflinks)
          • Quantify to reference (Partek E/M)
          • Quantify regions
          • HTSeq
          • Count feature barcodes
          • Salmon
        • Filtering
          • Filter features
          • Filter groups (samples or cells)
          • Filter barcodes
          • Split by attribute
          • Downsample Cells
        • Normalization and scaling
          • Impute low expression
          • Impute missing values
          • Normalization
          • Normalize to baseline
          • Normalize to housekeeping genes
          • Scran deconvolution
          • SCTransform
          • TF-IDF normalization
        • Batch removal
          • General linear model
          • Harmony
          • Seurat3 integration
        • Differential Analysis
          • GSA
          • ANOVA/LIMMA-trend/LIMMA-voom
          • Kruskal-Wallis
          • Detect alt-splicing (ANOVA)
          • DESeq2(R) vs DESeq2
          • Hurdle model
          • Compute biomarkers
          • Transcript Expression Analysis - Cuffdiff
          • Troubleshooting
        • Survival Analysis with Cox regression and Kaplan-Meier analysis - Partek Flow
        • Exploratory Analysis
          • Graph-based Clustering
          • K-means Clustering
          • Compare Clusters
          • PCA
          • t-SNE
          • UMAP
          • Hierarchical Clustering
          • AUCell
          • Find multimodal neighbors
          • SVD
          • CellPhoneDB
        • Trajectory Analysis
          • Trajectory Analysis (Monocle 2)
          • Trajectory Analysis (Monocle 3)
        • Variant Callers
          • SAMtools
          • FreeBayes
          • LoFreq
        • Variant Analysis
          • Fusion Gene Detection
          • Annotate Variants
          • Annotate Variants (SnpEff)
          • Annotate Variants (VEP)
          • Filter Variants
          • Summarize Cohort Mutations
          • Combine Variants
        • Copy Number Analysis (CNVkit)
        • Peak Callers (MACS2)
        • Peak analysis
          • Annotate Peaks
          • Filter peaks
          • Promoter sum matrix
        • Motif Detection
        • Metagenomics
          • Kraken
          • Alpha & beta diversity
          • Choose taxonomic level
        • 10x Genomics
          • Cell Ranger - Gene Expression
          • Cell Ranger - ATAC
          • Space Ranger
          • STARsolo
        • V(D)J Analysis
        • Biological Interpretation
          • Gene Set Enrichment
          • GSEA
        • Correlation
          • Correlation analysis
          • Sample Correlation
          • Similarity matrix
        • Export
        • Classification
        • Feature linkage analysis
      • Data Viewer
      • Visualizations
        • Chromosome View
          • Launching the Chromosome View
          • Navigating Through the View
          • Selecting Data Tracks for Visualization
          • Visualizing the Results Using Data Tracks
          • Annotating the Results
          • Customizing the View
        • Dot Plot
        • Volcano Plot
        • List Generator (Venn Diagram)
        • Sankey Plot
        • Transcription Start Site (TSS) Plot
        • Sources of variation plot
        • Interaction Plots
        • Correlation Plot
        • Pie Chart
        • Histograms
        • Heatmaps
        • PCA, UMAP and tSNE scatter plots
        • Stacked Violin Plot
      • Pipelines
        • Making a Pipeline
        • Running a Pipeline
        • Downloading and Sharing a Pipeline
        • Previewing a Pipeline
        • Deleting a Pipeline
        • Importing a Pipeline
      • Large File Viewer
      • Settings
        • Personal
          • My Profile
          • My Preferences
          • Forgot Password
        • System
          • System Information
          • System Preferences
          • LDAP Configuration
        • Components
          • Filter Management
          • Library File Management
            • Library File Management Settings
            • Library File Management Page
            • Selecting an Assembly
            • Library Files
            • Update Library Index
            • Creating an Assembly on the Library File Management Page
            • Adding Library Files on the Library File Management Page
            • Adding a Reference Sequence
            • Adding a Cytoband
            • Adding Reference Aligner Indexes
            • Adding a Gene Set
            • Adding a Variant Annotation Database
            • Adding a SnpEff Variant Database
            • Adding a Variant Effect Predictor (VEP) Database
            • Adding an Annotation Model
            • Adding Aligner Indexes Based on an Annotation Model
            • Adding Library Files from Within a Project
            • Microarray Library Files
            • Adding Prep kit
            • Removing Library Files
          • Option Set Management
          • Task Management
          • Pipeline managment
          • Lists
        • Access
          • User Management
          • Group Management
          • Licensing
          • Directory Permissions
          • Access Control Log
          • Failed Logins
          • Orphaned files
        • Usage
          • System Queue
          • System Resources
          • Usage Report
      • Server Management
        • Backing Up the Database
        • System Administrator Guide (Linux)
        • Diagnosing Issues
        • Moving Data
        • Partek Flow Worker Allocator
      • Enterprise Features and Toolkits
        • REST API
          • REST API Command List
      • Microarray Toolkit
        • Importing Custom Microarrays
      • Glossary
    • Webinars
    • Blog Posts
      • How to select the best single cell quality control thresholds
      • Cellular Differentiation Using Trajectory Analysis & Single Cell RNA-Seq Data
      • Spatial transcriptomics—what’s the big deal and why you should do it
      • Detecting differential gene expression in single cell RNA-Seq analysis
      • Batch remover for single cell data
      • How to perform single cell RNA sequencing: exploratory analysis
      • Single Cell Multiomics Analysis: Strategies for Integration
      • Pathway Analysis: ANOVA vs. Enrichment Analysis
      • Studying Immunotherapy with Multiomics: Simultaneous Measurement of Gene and Protein
      • How to Integrate ChIP-Seq and RNA-Seq Data
      • Enjoy Responsibly!
      • To Boldly Go…
      • Get to Know Your Cell
      • Aliens Among Us: How I Analyzed Non-Model Organism Data in Partek Flow
    • White Papers
      • Understanding Reads in RNA-Seq Analysis
      • RNA-Seq Quantification
      • Gene-specific Analysis
      • Gene Set ANOVA
      • Partek Flow Security
      • Single Cell Scaling
      • UMI Deduplication in Partek Flow
      • Mapping error statistics
    • Release Notes
      • Release Notes Archive - Partek Flow 10
  • Partek Genomics Suite
    • Installation Guide
      • Minimum System Requirements
      • Computer Host ID Retrieval
      • Node Locked Installation
        • Windows Installation
        • Macintosh Installation
      • Floating/Locked Floating Installation
        • Linux Installation
          • FlexNet Installation on Linux
        • Installing FlexNet on Windows
        • License Server FAQ's
        • Client Computer Connection to License Server
      • Uninstalling Partek Genomics Suite
      • Updating to Version 7.0
      • License Types
      • Installation FAQs
    • User Manual
      • Lists
        • Importing a text file list
        • Adding annotations to a gene list
        • Tasks available for a gene list
        • Starting with a list of genomic regions
        • Starting with a list of SNPs
        • Importing a BED file
        • Additional options for lists
      • Annotation
      • Hierarchical Clustering Analysis
      • Gene Ontology ANOVA
        • Implementation Details
        • Configuring the GO ANOVA Dialog
        • Performing GO ANOVA
        • GO ANOVA Output
        • GO ANOVA Visualisations
        • Recommended Filters
      • Visualizations
        • Dot Plot
        • Profile Plot
        • XY Plot / Bar Chart
        • Volcano Plot
        • Scatter Plot and MA Plot
        • Sort Rows by Prototype
        • Manhattan Plot
        • Violin Plot
      • Visualizing NGS Data
      • Chromosome View
      • Methylation Workflows
      • Trio/Duo Analysis
      • Association Analysis
      • LOH detection with an allele ratio spreadsheet
      • Import data from Agilent feature extraction software
      • Illumina GenomeStudio Plugin
        • Import gene expression data
        • Import Genotype Data
        • Export CNV data to Illumina GenomeStudio using Partek report plug-in
        • Import data from Illumina GenomeStudio using Partek plug-in
        • Export methylation data to Illumina GenomeStudio using Partek report plug-in
    • Tutorials
      • Gene Expression Analysis
        • Importing Affymetrix CEL files
        • Adding sample information
        • Exploring gene expression data
        • Identifying differentially expressed genes using ANOVA
        • Creating gene lists from ANOVA results
        • Performing hierarchical clustering
        • Adding gene annotations
      • Gene Expression Analysis with Batch Effects
        • Importing the data set
        • Adding an annotation link
        • Exploring the data set with PCA
        • Detect differentially expressed genes with ANOVA
        • Removing batch effects
        • Creating a gene list using the Venn Diagram
        • Hierarchical clustering using a gene list
        • GO enrichment using a gene list
      • Differential Methylation Analysis
        • Import and normalize methylation data
        • Annotate samples
        • Perform data quality analysis and quality control
        • Detect differentially methylated loci
        • Create a marker list
        • Filter loci with the interactive filter
        • Obtain methylation signatures
        • Visualize methylation at each locus
        • Perform gene set and pathway analysis
        • Detect differentially methylated CpG islands
        • Optional: Add UCSC CpG island annotations
        • Optional: Use MethylationEPIC for CNV analysis
        • Optional: Import a Partek Project from Genome Studio
      • Partek Pathway
        • Performing pathway enrichment
        • Analyzing pathway enrichment in Partek Genomics Suite
        • Analyzing pathway enrichment in Partek Pathway
      • Gene Ontology Enrichment
        • Open a zipped project
        • Perform GO enrichment analysis
      • RNA-Seq Analysis
        • Importing aligned reads
        • Adding sample attributes
        • RNA-Seq mRNA quantification
        • Detecting differential expression in RNA-Seq data
        • Creating a gene list with advanced options
        • Visualizing mapped reads with Chromosome View
        • Visualizing differential isoform expression
        • Gene Ontology (GO) Enrichment
        • Analyzing the unexplained regions spreadsheet
      • ChIP-Seq Analysis
        • Importing ChIP-Seq data
        • Quality control for ChIP-Seq samples
        • Detecting peaks and enriched regions in ChIP-Seq data
        • Creating a list of enriched regions
        • Identifying novel and known motifs
        • Finding nearest genomic features
        • Visualizing reads and enriched regions
      • Survival Analysis
        • Kaplan-Meier Survival Analysis
        • Cox Regression Analysis
      • Model Selection Tool
      • Copy Number Analysis
        • Importing Copy Number Data
        • Exploring the data with PCA
        • Creating Copy Number from Allele Intensities
        • Detecting regions with copy number variation
        • Creating a list of regions
        • Finding genes with copy number variation
        • Optional: Additional options for annotating regions
        • Optional: GC wave correction for Affymetrix CEL files
        • Optional: Integrating copy number with LOH and AsCN
      • Loss of Heterozygosity
      • Allele Specific Copy Number
      • Gene Expression - Aging Study
      • miRNA Expression and Integration with Gene Expression
        • Analyze differentially expressed miRNAs
        • Integrate miRNA and Gene Expression data
      • Promoter Tiling Array
      • Human Exon Array
        • Importing Human Exon Array
        • Gene-level Analysis of Exon Array
        • Alt-Splicing Analysis of Exon Array
      • NCBI GEO Importer
    • Webinars
    • White Papers
      • Allele Intensity Import
      • Allele-Specific Copy Number
      • Calculating Genotype Likelihoods
      • ChIP-Seq Peak Detection
      • Detect Regions of Significance
      • Genomic Segmentation
      • Loss of Heterozygosity Analysis
      • Motif Discovery Methods
      • Partek Genomics Suite Security
      • Reads in RNA-Seq
      • RNA-Seq Methods
      • Unpaired Copy Number Estimation
    • Release Notes
    • Version Updates
    • TeamViewer Instructions
  • Getting Help
    • TeamViewer Instructions
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On this page
  • Why did my import fail?
  • Why can’t I find the RPKM method on the normalization menu (I see FPKM)?
  • Why don't you have RPM in your normalization?
  • What is a canonical transcript?
  • Why are there decimal values in the Partek E/M quantification output?
  • Why can't I view PCA on Google Chrome and I am being pointed to WebGL instructions?
  • I have two projects that I ran separately. I want to combine the two projects without repeating the alignment step. How can I combine the aligned reads?
  • Why is the number of variants listed in the variant reports and summarize cohort mutations report different?
  • Additional Assistance
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  1. Partek Flow
  2. User Manual

Partek Flow FAQs

Last updated 7 months ago

Here are some Frequently Asked Questions about Partek Flow software based on real support calls.

Why did my import fail?

The most common form of failure is truncation. Ensure the transfer is complete before importing the data and that the file size and MD5 reported by Flow match the original data.

For bam files make sure the SAMtools utility can sort and index the file.

For fastq files run gunzip -c filename | tail and look for the message "unexpected end of file" or a missing component of the last read.

Why can’t I find the RPKM method on the normalization menu (I see FPKM)?

When working with paired data it should be the case that FPKM is available, and when working with single end data RPKM should be available. These metrics are essentially analogous, but based on the underlying method used for calculation (accounting for two reads mapping to 1 fragment and not counting twice for paired end data). Here is a simple description of the differences in calculation between RPKM and FPKM: .

Why don't you have RPM in your normalization?

RPM (reads per million) is the same as Total Count. Please use Total Count.

What is a canonical transcript?

For genes with multiple transcripts, one of the transcripts is picked as the canonical transcript. Based on the UCSC definition from the table browser,

knownCanonical - identifies the canonical isoform of each cluster ID, or gene. Generally, this is the longest isoform.

we define the canonical transcript as either the longest CDS (coding DNA sequence) if the gene has translated transcripts, or the longest cDNA.

Why are there decimal values in the Partek E/M quantification output?

The Partek E/M quantification algorithm can give decimal values because of multi-mapping reads (the same read potentially aligning to multiple locations) and overlapping transcripts/genes (a read that maps to a location with multiple transcripts or genes at that location). In these scenarios, the read count will be split.

For example, if a read maps to two potential locations, then that read contributes 0.5 counts to the first location and 0.5 counts to the second location. Similarly, if a read maps to one location with two overlapping genes, then that read contributes 0.5 counts to the first gene and 0.5 counts to the second gene.

If you need to remove the decimal points for downstream analysis outside of Partek Flow, you can round the values to the nearest integer.

Why can't I view PCA on Google Chrome and I am being pointed to WebGL instructions?

  • For Mac, this is the command: /Applications/Google\ Chrome.app/Contents/MacOS/Google\ Chrome --ignore-gpu-blacklist

  • Here is a video of a workaround and also creating an automated tool in a Mac (02:55):

https://github.com/illumina-swi/partek-docs/assets/167460925/d7a7ffc4-1b45-4e1b-b55c-644f3b05bb3e

I have two projects that I ran separately. I want to combine the two projects without repeating the alignment step. How can I combine the aligned reads?

Create a third project. Import two sets of BAM files.

Project 1 - In Data Tab, take note of your project output folder. Also note the aligner that you used.

Project 2 - In Data Tab, take note of your project output folder. Also note the aligner that you used.

Project 3 - Create project. Import Data > Automatically create samples from files > Find the project output folder of Project 1 (should be in Partek Flow server) > Find the folder for the aligner you used > Within that folder should be the BAM files > Select the BAM files > Create sample. Repeat same process for Project 2.

Now you will have all samples as one data node (aligned reads) in Project 3. You can proceed with downstream analysis (normalization, differential gene expression, etc).

Why is the number of variants listed in the variant reports and summarize cohort mutations report different?

For variants with multiple alternative alleles, the variant has one row for all alternative alleles, while the summarize cohort mutations report lists each alternative allele on a separate rows. The number of variants listed at the top of the each report is calculated from the number of rows in the report.

Additional Assistance

If you are using Google Chrome, some graphics card (particularly in older Mac laptops) are and they affect the ability to run WebGL (needed to display PCA). You may wish to use a different browser (Mozilla Firefox) or you can try running Google Chrome while ignoring any blacklisted hardware:

The app created in the video is available for download here (). Unzip and make sure to exit Google Chrome complete before running the app.

If you need additional assistance, please visit to submit a help ticket or find phone numbers for regional support.

http://www.rna-seqblog.com/rpkm-fpkm-and-tpm-clearly-explained
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