I am using QIIME2 for the first time and applying my samples after running the “Moving Pictures” tutorial.
During the “Moving Pictures” tutorial process using my sample, a problem occurred in the part of applying the metadata file.
My sample was NGS in the forward single strand (16S V3-V4 region) using Thermofisher’s Ion S5 equipment, and since there is no barcodes file, the raw data of Fastq was imported using the “Fastq manifest” format. After that, denoising was performed using DADA2. As the next step, in the process of the Feature Table, the metadata file for my sample was written directly, and an error occurred when applying it to qiime2.
It is an error with the following content.
There was an issue with loading the file sample-metadata_mov2.csv as metadata:
Found directive ‘#q2:types,categorical,categorical,categorical’ while searching for header. Directives may only appear immediately after the header.
There may be more errors present in the metadata file. To get a full report, sample/feature metadata files can be validated with Keemei: https://keemei.qiime2.org
Find details on QIIME 2 metadata requirements here: Metadata in QIIME 2 — QIIME 2 2020.2.0 documentation
The above error occurred continuously even though the contents of the metadata in the Qiime2 tutorial were checked and Keemei validated and confirmed that there was no problem in the metadata format.
The command in qiime2 proceeded to the following contents.
qiime feature-table summarize
The contents of the metadata file are as follows.
I want to know if there is a metadata error or another error.
Ask for advice.
#SampleID BarcodeSequence LinkerPrimerSequence Description
#q2:types categorical categorical categorical
sample-1 CTAAGGTAAC CCACTACGCCTCCGCTTTCCTCTCTATGGGCAGTCGGTGAT C11P1
sample-2 TAAGGAGAAC CCACTACGCCTCCGCTTTCCTCTCTATGGGCAGTCGGTGAT C11P2
sample-3 AAGAGGATTC CCACTACGCCTCCGCTTTCCTCTCTATGGGCAGTCGGTGAT C11P3
sample-4 TACCAAGATC CCACTACGCCTCCGCTTTCCTCTCTATGGGCAGTCGGTGAT C11P4
sample-5 CAGAAGGAAC CCACTACGCCTCCGCTTTCCTCTCTATGGGCAGTCGGTGAT C11P5
sample-6 CTGCAAGTTC CCACTACGCCTCCGCTTTCCTCTCTATGGGCAGTCGGTGAT C11P6
sample-7 TTCGTGATTC CCACTACGCCTCCGCTTTCCTCTCTATGGGCAGTCGGTGAT C11P7
sample-8 TTCCGATAAC CCACTACGCCTCCGCTTTCCTCTCTATGGGCAGTCGGTGAT C11P8
sample-9 TGAGCGGAAC CCACTACGCCTCCGCTTTCCTCTCTATGGGCAGTCGGTGAT C11P9
sample-10 CTGACCGAAC CCACTACGCCTCCGCTTTCCTCTCTATGGGCAGTCGGTGAT C11P10