Case Studies in Functional Genomics

Explore data analysis of several experimental protocols, using open source software, including R and Bioconductor.

Course Description

We will explain how to start with raw data, and perform the standard processing and normalization steps to get to the point where one can investigate relevant biological questions. Throughout the case studies, we will make use of exploratory plots to get a general overview of the shape of the data and the result of the experiment. We start with RNA-seq data analysis covering basic concepts of RNA-seq and a first look at FASTQ files. We will also go over quality control of FASTQ files; aligning RNA-seq reads; visualizing alignments and move on to analyzing RNA-seq at the gene-level: counting reads in genes; Exploratory Data Analysis and variance stabilization for counts; count-based differential expression; normalization and batch effects. Finally, we cover RNA-seq at the transcript-level: inferring expression of transcripts (i.e. a... Read More »

We will explain how to start with raw data, and perform the standard processing and normalization steps to get to the point where one can investigate relevant biological questions. Throughout the case studies, we will make use of exploratory plots to get a general overview of the shape of the data and the result of the experiment. We start with RNA-seq data analysis covering basic concepts of RNA-seq and a first look at FASTQ files. We will also go over quality control of FASTQ files; aligning RNA-seq reads; visualizing alignments and move on to analyzing RNA-seq at the gene-level: counting reads in genes; Exploratory Data Analysis and variance stabilization for counts; count-based differential expression; normalization and batch effects. Finally, we cover RNA-seq at the transcript-level: inferring expression of transcripts (i.e. alternative isoforms); differential exon usage. We will learn the basic steps in analyzing DNA methylation data, including reading the raw data, normalization, and finding regions of differential methylation across multiple samples. The course will end with a brief description of the basic steps for analyzing ChIP-seq datasets, from read alignment, to peak calling, and assessing differential binding patterns across multiple samples.

Given the diversity in educational background of our students we have divided the series into seven parts. You can take the entire series or individual courses that interest you. If you are a statistician you should consider skipping the first two or three courses, similarly, if you are biologists you should consider skipping some of the introductory biology lectures. Note that the statistics and programming aspects of the class ramp up in difficulty relatively quickly across the first three courses. By the third course will be teaching advanced statistical concepts such as hierarchical models and by the fourth advanced software engineering skills, such as parallel computing and reproducible research concepts.


This class was supported in part by NIH grant R25GM114818.

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Course Outcomes:
  • Mapping reads
  • Quality assessment of Next Generation Data
  • Analyzing RNA-seq data
  • Analyzing DNA methylation data
  • Analyzing ChIP Seq data
Course Details:

Prerequisites

  • PH525.3x: Statistical Inference and Modeling for High-throughput Experiments
  • PH525.4x: High-Dimensional Data Analysis
  • About Instructor:

    Rafael Irizarry - Professor of Biostatistics

    Rafael Irizarry is a Professor of Biostatistics at the Harvard T.H. Chan School of Public Health and a Professor of Biostatistics and Computational Biology at the Dana Farber Cancer Institute. For the past 15 years, Dr. Irizarry’s research has focused on the analysis of genomics data. During this time, he has also has taught several classes, all related to applied statistics. Dr. Irizarry is one of the founders of the Bioconductor Project, an open source and open development software project for the analysis of genomic data. His publications related to these topics have been highly cited and his software implementations widely downloaded.


    Michael Love - Assistant Professor, Departments of Biostatistics and Genetics

    Michael Love is Assistant Professor in the Departments of Biostatistics and Genetics at UNC Gillings School of Global Public Health. Dr. Love received his bachelor’s in mathematics in 2005 from Stanford University, his master’s in statistics in 2010 from Stanford University, and his Ph.D. in Computational Biology in 2013 from the Freie Universität Berlin. Dr. Love uses statistical models to infer biologically meaningful patterns from high-throughput sequencing data, and develops open-source statistical software for the Bioconductor Project.


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