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. 2012 Nov:Chapter 1:Unit 1E.5..
doi: 10.1002/9780471729259.mc01e05s27.

Using QIIME to analyze 16S rRNA gene sequences from microbial communities

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Using QIIME to analyze 16S rRNA gene sequences from microbial communities

Justin Kuczynski et al. Curr Protoc Microbiol. 2012 Nov.

Abstract

QIIME (canonically pronounced "chime") is a software application that performs microbial community analysis. It is an acronym for Quantitative Insights Into Microbial Ecology, and has been used to analyze and interpret nucleic acid sequence data from fungal, viral, bacterial, and archaeal communities. The following protocols describe how to install QIIME on a single computer and use it to analyze microbial 16S sequence data from nine distinct microbial communities.

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Figures

Figure 1E.5.1
Figure 1E.5.1
A screenshot of the QIIME VirtualBox, with the terminal icon indicated, and a terminal window open.
Figure 1E.5.2
Figure 1E.5.2
Contents of the mapping file ( Fasting_Map.txt). Note that the SampleIDs contain only letters, numbers, and period characters.
Figure 1E.5.3
Figure 1E.5.3
The first few lines of the taxonomy assignment file, showing on each line the OTU identifier, the representative sequence identifier, the taxonomy assigned to that sequence, and the confidence in that assignment.
Figure 1E.5.4
Figure 1E.5.4
A visualization of the phylogenetic tree using FigTree. The tips are unlabeled here, but can be inspected interactively.
Figure 1E.5.5
Figure 1E.5.5
An OTU table heatmap, showing the relative abundance of each OTU within each microbial community.
Figure 1E.5.6
Figure 1E.5.6
Magic-Table visualization of the OTU table heatmap.
Figure 1E.5.7
Figure 1E.5.7
An OTU table heatmap showing taxonomy assignment for each OTU.
Figure 1E.5.8
Figure 1E.5.8
An area chart showing the relative abundance of each phylum within each microbial community.
Figure 1E.5.9
Figure 1E.5.9
A bar chart of phylum level abundance within communities, similar to Figure 10.7.8.
Figure 1E.5.10
Figure 1E.5.10
A Web browser window displaying rarefaction plots. The vertical axis displays the diversity of the community, while the horizontal axis displays the number of sequences considered in the diversity calculation. Each line on the figure represents the average of all microbial belonging to a group within a category: here the green line represents all fasted mouse communities, and the blue line represents the control communities.
Figure 1E.5.11
Figure 1E.5.11
A visualization of bootstrap-supported hierarchical clustering of the 9 microbial communities under investigation. Note that the fasted mouse communities ( PC.6××) cluster together, and the result is supported by jackknife tests (red implies > 75% support).
Figure 1E.5.12
Figure 1E.5.12
A Principal Coordinates plot of the 9 communities, showing jackknife-supported confidence ellipsoids. The first two principal axes are shown.

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