Erroe while running alpha diversity

Hi,

I am facing problem with the command given below. Also, find the attached metadata file. Can you please let me know what is the problem in my metadata file or command?

Thank you

(qiime2-2021.8) [email protected] empsingle % qiime diversity alpha-group-significance --i-alpha-diversity core-metrics-results/faith_pd_vector.qza --m-metadata-file metadata.tsv --o-visualization visualizations/alpha_diversity/faith_pd.qzvmetadata.tsv (180 Bytes)

Plugin error from diversity:

Metadata does not contain any columns that satisfy this visualizer's requirements. There must be at least one metadata column that contains categorical data, isn't empty, doesn't consist of unique values, and doesn't consist of exactly one value.

Debug info has been saved to /var/folders/pq/2rvx277j7vq8z1tkbjypgzww0000gp/T/qiime2-q2cli-err-3jks6h53.log

Hello! Actually, the error is self explanatory.

So, the plugin is complaining that your metadata file do not have any categorical columns that contain non-unique values. In other words, you need columns that will divide your dataset into several groups and plugin can perform stat analysis between them. For example, it can be the source of the samples, sampling sites, treatments and so on. It is important that each group should be represented with enough replicates.

Hi @timanix ,

Thank you for your response, I changed my metadata file, but I received the same error given below

(qiime2-2021.8) [email protected] empsingle % qiime diversity alpha-group-significance --i-alpha-diversity core-metrics-results/faith_pd_vector.qza --m-metadata-file metadata.tsv --o-visualization visualizations/alpha_diversity/faith_pd.qzv

Plugin error from diversity:

Metadata does not contain any columns that satisfy this visualizer's requirements. There must be at least one metadata column that contains categorical data, isn't empty, doesn't consist of unique values, and doesn't consist of exactly one value.

Debug info has been saved to /var/folders/pq/2rvx277j7vq8z1tkbjypgzww0000gp/T/qiime2-q2cli-err-igsawsvv.logmetadata.tsv (206 Bytes)

Hello again,

As I already wrote, your dataset should include replicates for each group. In the metadata you shared each sample is unique in the categorical column. And 2 samples is not enough for stat. analysis. So you need more samples in your dataset.

Hi @timanix ,

Thank you for your help. It worked with other file that has enough dataset.

With regards,
Aditya