Frequently Asked Questions¶
What kind of data can I upload to Qiita for processing?¶
Processing in Qiita requires 3 things: raw data, sample and prep information files. Here you can find a list of currently supported raw files files. Note that we are accepting any kind of target gene (16S, 18S, ITS, whatever). You can also upload WGS however, WGS processing is not ready.
What’s the difference between a sample and a prep information file?¶
A sample information file describes the samples in a study, including environmental factors relating to the associated host. The prep information file has information on how the sample was processed in the wet lab. If you collected 100 samples for your study, you will need 100 rows in your sample information file describing each of them, and additional rows for blanks and other control samples. If you prepared 95 of them for 16S and 50 of them for 18S, you will need 2 prep information files: one with 95 rows describing the preparation for 16S, and another one with 50 describing the 18S. For a more complex example go here and for examples of these files you can go to the “Upload instructions” here.
Example study processing workflow¶
A few more instructions: for the example above the workflow should be:
- Create a new study.
- Add a sample information file. You can add 1, try to process it and the system will let you know if you have errors or missing columns. The most common errors are: the sample name column should be named sample_name, duplicated sample names are not permitted. For a full list of required fields, visit Getting Started Guide.
- Add a prep information file to your study for each data type. The prep information file should contain all the samples in the sample information file or a subset. If you have more than one FASTQ file set (forward, reverse (optional) and barcodes) you will need to add a run_prefix column, see Prepare information files. A prep information file and a QIIME compatible mapping file will be available for download after the prep information file is added successfully.
- Upload and link your raw data to each of your prep information files. Depending on your barcoding/sequencing strategy you might need 1 or more raw data file sets. If you have 2 raw data sets you may have to rename one set so that each set has a different name. If they have the same name they will over-write on upload. Note that you can have one FASTQ file set linked to more than one prep information file.
- Preprocess your files. For target gene amplicon sequencing, this will demux and QC. There are multiple options for preprocessing depending on the barcode format and the data output from the sequencing center - this may require a series of trial and error to establish the correct option for your data files. After demultiplexing a log file is generated with statistics about the files demultiplexed including the number of sequences assigned per sample.
- Process each of your preprocessed data types. For target gene, this will perform closed OTU picking against the latest version of Greengenes and can be quite time consuming depending on the number of samples and the depth of sequencing.
How to solve unzip errors?¶
When downloading large zip files within Qiita there is a change that you will get an error like: “start of central directory not found; zipfile corrupt”. This issue arises from using old versions of zip and you need to have unzip >= 6.0.0. To check you unzip version you can run: unzip -v.
To update your unzip for most operating systems you can simply use your regular package admin program. However, for Mac we suggest using this version of unzip.