
A brief introduction into what the course is and what Next Generation Sequencing is in relation to RNA sequencing.
Learn how to obtain the needed files to download the RNA sequencing reads onto Galaxy, bypassing the need to limit yourself by your own network speed or storage limitations.
Learn how to upload files onto Galaxy and use the SRA tool to download the needed sequencing files from the SRA database.
Learn how to quality control next-generation sequencing data using FastQC and compile reports using MulitQC
Learn how to get the files you need from ENSEMBL.
Learn how to use STAR for RNA sequencing read alignment to a particular reference genome.
Use infer experiment to figure out details of the RNA sequencing experiment that are vital for analysis but may be left out of publication material.
Learn how to use FeatureCounts a count table of the STAR alignment and the exon and gene level.
Learn how to carry out differential expression analysis using DESeq2 and how to interpret the results once a comparison is made.
Learn how to annotate the DESeq2 output with your gtf file and prepare a file for the GOseq.
Learn how to interpret the GOseq results
Learn how to edit the table generated from DESeq2 to allow input into Pathview and also where to find the KEGG pathway lists.
Learn how to interpret the KEGG pathway results and also what useful information that these can tell you.
Learn how to use DEGUST to carry out a differential gene expression analysis without the need for Galaxy. You will also learn how to interpret the many graphs that DEGUST can create.
Ever wonder which technologies allow researchers to discover new markers of cancer or to get a greater understanding of genetic diseases? Or even just what genes are important for cellular growth?
This is usually carried out using an application of Next Generation Sequencing Technology called RNA sequencing. Throughout this course, you will be equipped with the tools and knowledge to not only understand but perform RNA sequencing and discover how the transcriptome of a cell changes throughout its growth cycle. To avoid the need for complex software installations, coding experience and in some cases a Linux operating system we will be using a free bioinformatics tool called Galaxy for the whole analysis! Not only that, but we will also be using the STAR pipeline which is currently supported by the ENCODE project!
Once you've completed this course you will know how to:
Download publically available data from papers straight onto Galaxy.
Obtain the needed raw files for genome alignment.
Perform genome alignment using a tool called STAR.
Create count tables from your alignment using FeatureCounts.
Carry out a differential expression using DESeq2 to find out what changes between a cell on day 4 Vs day 7 of growth.
Carry out gene ontology analysis to understand what pathways are up and down-regulated.
Use Pathview to create annotated KEGG maps that can be used to look at specific pathways in more detail.
Use a web browser-based tool called DEGUST as an alternative to using DESeq2.
Practical Based
The course has one initial lecture explaining some of the basics of sequencing and what RNA sequencing can be used for. Then it's straight into the practical! Throughout the 14 lectures, you are guided step by step through the process from downloading the data to how you could potentially interpret the data at the final stages. Unlike most courses, the process is not simplistic. The project has real-world issues, such as dealing with galaxies limitations and how you can get around them with some initiative!
This course is made for anyone that has an interest in Next-Generation Sequencing and the technologies currently being used to make breakthroughs in genetic and medical research! The course is also meant for beginners in RNA-seq to learn the general process and complete a full walkthrough that is applicable to there own data!