Difference between revisions of "Amplicon sequencing"
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Revision as of 13:58, 24 August 2015
Amplicon sequencing and downstream metagenetic analysis
Often, Illumina amplicon sequencing is used to assess the diversity and composition of microbial populations. For many purposes, the entire downstream bioinformatic analysis may be carried out using only the Qiime pipeline. However, for unusual or very large data, some steps will not work to satisfaction. Hence, alternative bioinformatic programs may have to be found.
This wiki contains a quick overview over the typical steps included in the analysis of amplicon sequencing data. Also, some of the alternative programs are listed, together with reasons to use them, rather than Qiime.
If using Qiime, consult the documentation at http://qiime.org . Qiime is installed on Abel; the default version is v1.5.0, but versions v1.8.0 and v1.9.1 are available as well. Load the default version of Qiime as
module load qiime
or the newest verion as
module load qiime/1.9.1
Quality control of fastq read files
-Removal of PCR primer sequences
-Converting fastq files into fasta files
-De-replication of reads
-Clustering of reads
-Removing singletons
-Selecting representative sequences for clusters
-Chimera-checking
-Assigning taxonomical identifiers
-Analysing OTU tables
