Contents ◾ ix
Chapter 6 ◾ Chromatin Immunoprecipitation Sequencing
213
6.1 INTRODUCTION TO CHROMATIN IMMUNOPRECIPITATION
213
6.2 CHIP SEQUENCING
214
6.3 CHIP-SEQ ANALYSIS WORKFLOW
215
6.3.1
Downloading the Raw Data
217
6.3.2
Quality Control
218
6.3.3
ChIP-Seq and Input Read Mapping
219
6.3.4
ChIP-Seq Peak Calling with MACS3
223
6.3.5
Visualizing ChIP-Seq Enrichment in Genome Browser
226
6.3.6
Visualizing Peaks Distribution
229
6.3.6.1 ChIP-Seq Peaks’ Coverage Plot
230
6.3.6.2 Distribution of Peaks in Transcription Start Site (TSS)
Regions
233
6.3.6.3 Profile of Peaks along Gene Regions
234
6.3.7
Peak Annotation
235
6.3.7.1 Writing Annotations to Files
237
6.3.8
ChIP-Seq Functional Analysis
239
6.3.9
Motif Discovery
243
6.4 SUMMARY
250
REFERENCES
251
Chapter 7 ◾ Targeted Gene Metagenomic Data Analysis
253
7.1. INTRODUCTION TO METAGENOMICS
253
7.2 ANALYSIS WORKFLOW
254
7.2.1
Raw Data Preprocessing
254
7.2.2
Metagenomic Features
255
7.2.2.1 Clustering
255
7.2.2.2 Denoising
256
7.2.3
Taxonomy Assignment
258
7.2.3.1 Basic Local Alignment Search Tool
258
7.2.3.2 VSEARCH
259
7.2.3.3 Ribosomal Database Project
259
7.2.4
Construction of Phylogenetic Trees
260
7.2.5
Microbial Diversity Analysis
261
7.2.5.1 Alpha Diversity Indices
262
7.2.5.2 Beta Diversity
262