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Processing recommendations and pipelines #2049
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| dev/index.rst | ||
| faq.rst | ||
| resources.rst | ||
| processing-recommendations.rst | ||
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qiita_pet/support_files/doc/source/processing-recommendations.rst
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| Processing recommendations | ||
| ========================== | ||
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| Currently, Qiita supports the processing :sup:`(*)` of raw data from: | ||
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| #. Target gene barcoded sequencing | ||
| #. Shotgun sequencing | ||
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| Note that the selected processing are mainly guided so we can perform meta-analyses, this is combine different studies, even from different wet lab techniques or | ||
| sequencing technologies. Remember to check the :ref:`example_study_processing_workflow` before continuing. | ||
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| For more information about meta-analysis, examples and things to consider: | ||
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| - `"Tiny microbes, enormous impacts: what matters in gut microbiome studies?" <https://genomebiology.biomedcentral.com/articles/10.1186/s13059-016-1086-x>`_ | ||
| - `"Meta-analyses of studies of the human microbiota" <http://genome.cshlp.org/content/23/10/1704.short>`_. | ||
| - `"A Guide to Enterotypes across the Human Body: Meta-Analysis of Microbial Community Structures in Human Microbiome Datasets" <http://journals.plos.org/ploscompbiol/article?id=10.1371/journal.pcbi.1002863>`_. | ||
| - `"Dynamic changes in short- and long-term bacterial composition following fecal microbiota transplantation for recurrent Clostridium difficile infection" <http://microbiomejournal.biomedcentral.com/articles/10.1186/s40168-015-0070-0>`_. | ||
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| :sup:`(*)` Remember that you can also upload BIOM tables for plotting but not covered here because this is only for raw data. | ||
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| Target gene barcoded sequencing | ||
| ------------------------------- | ||
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| For this you can start with raw, not demultiplexed data or per_sample_FASTQ, see :ref:`example_study_processing_workflow`. Either way, you will need to | ||
| "Split libraries and QC", which uses the default in QIIME 1.9.1. Once your demultiplexed and QCed artifact is created you need to select which processing to perform. | ||
| There are two main ideologies/methodologies to process target gene data: sequence clustering and sequence cleanup. | ||
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| Sequencing cleanup (preferred) | ||
| ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ | ||
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| For this we use `deblur <https://github.com/biocore/deblur>`_. Here 2 BIOM tables are generated by default: fina.biom and final.only-16s.biom. The former is the full biom table, which can be used with any target gene and wetlab work; | ||
| the latter is the trimmed version to those sequences that match Greengenes at 80% similarity, a really basic and naive filtering. Each of those BIOM tables, is accompanied by a FASTA that contains | ||
| the representative sequences. The OTU IDs are given by the unique sequence. | ||
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| Note that deblur needs all sequences to be trimmed at the same length, thus the recommended pipeline is to trim everything at 150bp and the deblur. | ||
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| Sequencing clustering | ||
| ^^^^^^^^^^^^^^^^^^^^^ | ||
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| Here we use close reference picking, for an explanation of the different picking methods see | ||
| `"Subsampled open-reference clustering creates consistent, comprehensive OTU definitions and scales to billions of sequences" <https://peerj.com/articles/545/>`_. | ||
| Here we generate a single BIOM table with the OTUs/per-sample. The OTU IDs are given based on the reference database selected. | ||
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| Currently, we have the reference databases: Greengenes version 3_8-97, Silva 119 and Unite 7. Depending on your selection is if the reference has a phylogenetic tree. | ||
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| Shotgun sequencing | ||
| ------------------ | ||
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| Here you need to start with per_sample_FASTQ, we recommend to only upload already QC-ed and adaptor and human sequences removed FASTQ files. However, we have a step for | ||
| this preprocessing available in Qiita via `KneadData <https://bitbucket.org/biobakery/kneaddata/wiki/Home>`_. | ||
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| The recommended processing steps are: | ||
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| #. Remove adapters and human sequences from your files using KneadData. We currently have TruSeq3-PE-2 and NexteraPE-PE adaptor removal. | ||
| #. Use `HUMAnN2 <https://bitbucket.org/biobakery/humann2/wiki/Home>`_ to generate BIOM tables. | ||
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| For more information visit the `Shotgun Qiita Plugin GitHub page<https://github.com/qiita-spots/qp-shotgun>`. | ||
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I don't think we have a way to upload pre-QC'd data at the moment...