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.gmt
andh.all.v2024.1.Hs.symbols.gmt
are 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.