Hi
So I performed a couple of your recommended filtering steps - first chimera filtering and thencontingency based filtering for singletons for both my feature table and my representative sequences (code below). I did not perform quality filtering - I am working with 454 data, and I do not have fastq files.
Input:
-
filter feature table
qiime feature-table filter-features --i-table table-nonchimeric.qza --p-min-samples 2 --o-filtered-table table-nonchimeric-gt2.qza -
Summarize feature table
qiime feature-table summarize --i-table split-library-output/uchime-dn-out table-nonchimeric-gt2.qza
–o-visualization table-nonchimeric-gt2.qzv --m-sample-metadata-file sample-metadata.tsv * Number of features = 16529 -
Use filtered feature table to filter rep-seqs
qiime feature-table filter-seqs --i-data rep-seqs-nonchimeric.qza --i-table table-nonchimeric-gt2.qza --o-filtered-data rep-seqs-nonchimeric-gt2.qza -
Align using MAFFT (worked fine this time)
qiime alignment mafft --i-sequences rep-seqs-nonchimeric-gt2.qza --o-alignment rep-seqs-nonchimeric-gt2-aligned.qza -
Mask alignment
qiime alignment mask --i-alignment rep-seqs-nonchimeric-gt2-aligned.qza --o-masked-alignment masked-rep-seqs-nonchimeric-gt2-aligned.qza
-Make the tree
qiime phylogeny fasttree --i-alignment masked-rep-seqs-nonchimeric-gt2-aligned.qza --o-tree unrooted-tree2.qza
-Root the tree
qiime phylogeny midpoint-root --i-tree unrooted-tree2.qza --o-rooted-tree rooted-tree2.qza
- Perform diversity analyses
qiime diversity core-metrics-phylogenetic --i-phylogeny split_library_output/uchime-dn-out/rooted-tree.qza --i-table split_library_output/uchime-dn-out/table-nonchimeric-gt2.qza --p-sampling-depth 900 --m-metadata-file map_file.tsv --output-dir core-metrics-results-gt2
However, when I run this command I get a similar error to this post Diversity plug-in error where my feature IDs are again missing. I have checked the output table, rep-seqs file, and the tree, and the “missing” features are all present.
Thank you
Laura