Class Overview
This class introduces participants to bulk RNA sequencing analysis using the point-and-click software Partek Flow. Partek Flow enables researchers to build comprehensive workflows for analyzing data derived from many sequencing modalities on the bulk and single cell level (ie. RNA, ChIP/ATAC, CITE, and spatial transcriptomics). NCI holds an institutional license for this package. Please see https://bioinformatics.ccr.cancer.gov/docs/getting-started-with-partek-flow/ to learn how to get access and about different methods for transferring data to the NIH Partek Flow server. NHGRI also holds an institutional license to this software (see https://research.nhgri.nih.gov/bi-training.shtml). Scientists not affiliated with NCI or NHGRI can inquire with the NIH Library (https://www.nihlibrary.nih.gov/resources/tools/partek-flow).
Class Expectations / Learning Objectives
This class will not turn participants into expert bioinformaticians or Partek Flow users. However, concepts learned can be applied to other sequencing type using Partek Flow and provides a foundation for continual learning.
After this class, participants will become familiar with steps for analyzing bulk RNA sequencing data using Partek Flow, including:
- Creating new analysis project and importing FASTQ files into an analysis project.
- Performing quality control and cleanup of FASTQ files for use with downstream analyses.
- Aligning sequences in FASTQ files to reference genome and interpret alignment quality metrics.
- Generating gene expression table from aligned reads.
- Performing differential gene expression analysis.
- Constructing plots such as PCA, heatmap, and volcano plot to visualize RNA sequencing data.
Required Course Materials
This class is not hands-on. Experience using or access to Partek Flow is not required for participation.
Tip
For a review or introduction to RNA sequencing, see An Introduction to RNA-Seq: Overview of Expression Data Analysis.
Self Learning Material
- BTEP Getting Started with Partek Flow at NIH
- Check out the BTEP Video Archive for recordings of previous Partek Flow trainings
- Information regarding Partek Flow on Biowulf
- Partek Flow documentations
Contacts for Help
- BTEP: ncibtep@nih.gov
- Biowulf: staff@hpc.nih.gov
- Partek: support@partek.com
Download class data and try on your own
Click here to download the class data as a zip files to local computer. Macs should automatically unzip upon download but Windows users will have to unzip after download. Follow the instructions at https://bioinformatics.ccr.cancer.gov/docs/getting-started-with-partek-flow/data_transfer_pf_web/ to learn how to transfer data from personal computer to the NIH Partek Flow server.
The content in the zip file are:
22.fa: human chromosome 22 genome (source: Biostar Handbook: RNA-SEQ by Example, December 2021)22.gtf: human chromosome 22 gtf annotation (source: Biostar Handbook: RNA-SEQ by Example, December 2021)c6.all.v2024.1.Hs.symbols.gmtandh.all.v2024.1.Hs.symbols.gmtare the C6 and hallmark gene sets from msigdbhcc1395_fastqs: paired end fastq files for hcc1395 (source: Griffith lab RNA bio) - renaming of the files obtained from this source was required to fit the formats for some Partek Flow outputs (ie. tables and visualizations)
Tip
Upload 22.fa, 22.gtf, and the MSigDB gene sets to the user's Partek Flow library files. See https://documentation.partek.com/display/FLOWDOC/Library+File+Management to learn how.