Error: Failed to perform normalization! How to solve?

Hi developers:
Thanks for your effort in developing and maintaining this robust website.
I met an abnormal condition in processing the input ASV table in the Data Normalization module.
“Error: Failed to perform normalization!”
I chose the default settings in the Data Filter module, and I chose the “Rarefy to the minimum library size”, “Do not scale my data”, and “Do not transform my data” in the Data Normalization module.
So, how can I check the error details, or how to solve this?
Thanks for your attention and looking forward to your reply!

ASVs_table.txt (286.4 KB)
METADATA_grouped_by_site.txt (77 Bytes)
ASVs_tax.txt (698.3 KB)
The input files are attached.


Thanks for reporting the issue. It is caused by the unexpected symbols in your taxonomy table:

You can remove them or replace with correct format.

Hope this helps.

Thanks for your kindly reply in this OmicsForum and E-mail!
The processing can work on my data after correct the tax names.
Thanks a lot!
Best wishes!

I have faced same problem (failed to perform normalization), please help me out in this. i have attached the file on which i tried to do the analysis.

newgrouptaxonomy1.txt (17.2 KB)
newgroupmd.txt (372 Bytes)
newgroupFT.txt (6.3 KB)


I have checked your data. After the filtering step, there was only 3 features left. To get biological meaningful results, our website will keep the features that exist in at least 10% of the samples which will filter out most of your features. Please double check your abundance table. We will also consider this in our processing step if there are more similar cases.

Hope this helps.

what exactly should i look into the abundance table? And what if i do alter in low count filter and low variance filter? does it make any sense in positive or negative way?

Statistical analysis requires replicates (repeated observations) to estimate model parameters. If you open your data file, it contains only six samples each containing too many zeroes … most features appear only once. Most statistics will fail for such sparse data. If possible, increase sequencing depth or include more replicates (better both) when plan your next studies.

Our tools are designed to make robust or valid statistical analysis tasks easier. You can try different filters to see you can “trick” the system. However, you need to feel confident with the conclusion.

Hi there,

Thanks for your effort in developing and maintaining this comprehensive web-based platform,
I had the same issue "Error: Failed to perform normalization”. I chose the default settings in the Data Normalization, and I chose “Do not scale my data” - data rarefying, “Do not transform my data” - data transformation, and “Total sum scaling (TSS)” - data scaling. My ASVs data is normalized so I chose “Normalised data” in the data upload section. The input files are attached herewith, look forward to hearing your advice, many thanks,

Best regards,

asv_table.txt (196.8 KB)
taxa_table.txt (220.4 KB)
metadata.csv (363 Bytes)


I have the same problem (faild to perform normalization), please help me!
I attach the file in which I tried to do the analysis, my problem is only with my fungal data, with my bacteria data I have no problem,

Fungi.csv (64.2 KB)
metadata.csv (473 Bytes)

1 Like

Hi all, the issue has been fixed now. You can try again and let me know if you have more questions.

@Binod Your data contain very few features after filtration which may cause the failure of normalization. You can change the default filtration condition to include more features while still be biological meaningful.

1 Like

Thank you so much for checking my data, if I specify a minimum count “0” then the data is normalized but analysis such as alpha diversity, greengenes, stacked bar/area plot, etc. won’t work, this might be due to normalized input. look forward to hearing your advice, many thanks,

Best regards,