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DE Visualization
Navigate to the correct directory and then launch R:
cd $RNA_HOME/de/ballgown/ref_only
R
A separate R tutorial file has been provided in the github repo for part 2 of the tutorial: Tutorial_Part2_ballgown.R. Run the R commands detailed in the R script. All results are directed to pdf file(s). The output pdf files can be viewed in your browser at the following urls. Note, you must replace YOUR_IP_ADDRESS with your own amazon instance IP (e.g., 101.0.1.101)).
- http://YOUR_IP_ADDRESS/rnaseq/de/ballgown/ref_only/Tutorial_Part2_ballgown_output.pdf
Occasionally you may wish to reformat and work with stringtie output in R manually. Therefore we provide an optional/advanced tutorial on how to format your results for R and perform "old school" (non-ballgown analysis) on your data.
In this tutorial you will:
- Learn basic R usage and commands (common plots, and data manipulation tasks)
- Examine the expression estimates
- Create an MDS plot to visualize the differences between/among replicates, library prep methods and UHR versus HBR
- Examine the differential expression estimates
- Visualize the expression estimates and highlight those genes that appear to be differentially expressed
- Generate a list of the top differentially expressed genes
- Ask how reproducible technical replicates are.
Expression and differential expression files will be read into R. The R analysis will make use of the transcript-level expression and differential expression files from stringtie/ballgown. Navigate to the correct directory and then launch R:
cd $RNA_HOME/de/ballgown/ref_only/
R
A separate R file has been provided in the github repo for part 3 of the tutorial: Tutorial_Supplementary_R.R. Run the R commands detailed in the R script above.
The output file can be viewed in your browser at the following url. Note, you must replace YOUR_IP_ADDRESS with your own amazon instance IP (e.g., 101.0.1.101)).
- http://YOUR_IP_ADDRESS/rnaseq/de/ballgown/ref_only/Tutorial_Part3_Supplementary_R_output.pdf
Previous Section | This Section | Next Section |
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Differential Expression | DE Visualization | Kallisto |
NOTICE: This resource has been moved to rnabio.org. The version here will be maintained for legacy use only. All future development and maintenance will occur only at rnabio.org. Please proceed to rnabio.org for the current version of this course.
Table of Contents
Module 0: Authors | Citation | Syntax | Intro to AWS | Log into AWS | Unix | Environment | Resources
Module 1: Installation | Reference Genomes | Annotations | Indexing | Data | Data QC
Module 2: Adapter Trim | Alignment | IGV | Alignment Visualization | Alignment QC
Module 3: Expression | Differential Expression | DE Visualization
Module 4: Alignment Free - Kallisto
Module 5: Ref Guided | De novo | Merging | Differential Splicing | Splicing Visualization
Module 6: Trinity
Module 7: Trinotate
Appendix: Saving Results | Abbreviations | Lectures | Practical Exercise Solutions | Integrated Assignment | Proposed Improvements | AWS Setup