Tutorials

Tutorials are divided into two sections:

  • The Cookbook contains step-by-step instructions for performing common tasks, with detailed explanations of each step

  • In-depth includes longer discussions of issues that arise in high-throughput sequencing and genomics, and includes code examples when appropriate.

We suggest downloading the Demo dataset and following along.

Cookbook

Tutorial

Contents

Setting up a genome for analysis

Set up a genome for downstream analysis, using plastid or other toolkits

A simple alignment and quantitation workflow

Simplified workflow of how to align sequencing data

Arrays of counts at each transcript position

Retrieve a vector of high-throughput sequencing counts at each position in a transcript

Manipulating feature sequences

Fetch the sequences of regions of interest (e.g. transcripts) from a genome

Gene expression & translation efficiency

Compute gene expression measurements and translation efficicency using RNA-seq & ribosome profiling, and prepare data for differential expression analysis

Excluding (masking) regions of the genome

Exclude specific regions – for example, repetitive genome sequence that gives rise to multimapping reads – from analysis. Discussion of mask files.

Creating custom BED, BigBed, and GTF2 annotation files

Make a BED, BigBed, GTF2, or GFF3 file containing custom features.

Metagene analysis

Perform metagene analysis, using ribosome profiling data at the start codon as an example. Then, develop metagene analysis around a custom landmark for use with other data types

Ribosomal P-site offsets

Determine a P-site offset from ribosome profiling data

Read phasing in ribosome profiling

Estimate read phasing (triplet periodicity) of ribosome profiling data

Plotting tools

Demos of plotting tools in plastid.plotting

In-depth

Tutorial

Contents

Categories and formats of genomics data

Introduction & discussion to the types of data used in genomics, and the advantages and disadvantages of their various file formats

Coordinate systems used in genomics

Primer on the various coordinate systems used in genomics

Ambiguous read alignments

Issues arising when and strategies for handling multimapping reads

Read mapping functions

In-depth discussion of mapping rules, with code examples of how to write your own mapping rule for your own sequencing data type