For studying Qiime2, I have forward.fastq.gz, reverse.fastq.gz, barcodes.fastq.gz and mapping_file.tsv downloaded from Qiita (ID 11635). I think it matches the type EMP-paired and it looks like that there are no barocdes and primers in forward.fastq.gz and reverse.fastq.gz.
Here are my operation steps and error reporting
Step1: I have imported my files as EMP-paired using the command:
qiime tools import
Step2: I used the following code to demultiplex and got the "demux03.qza" successfully.
qiime demux emp-paired
qiime demux summarize
However, when viewing "demux-full.qzv", the sequence counts dropped dramatically after the first two samples and looked so strange.
Thanks for your reply,
I have checked the counts provided by Qiita, and they are truly more than that in our outcomes.
Also, I compared the number of lines contained in barcodes.fastq.gz, forward.fastq.gz and reverse.fastq.gz. The three raw files have the same number of lines.
I am not sure if I get what you mean. Could you tell me what should I do next?
Anxious anticipates your reply.
Thank you very much for your time and explanation.
The problem has been well solved when I rerun using --p-rev-comp-barcodes and --p-rev-comp-mapping-barcodes.
However, I still have a confusion. Could you please tell me how can I know whether there have rev_comp_barcode and rev_comp_mapping_barcodes as shown in your reply.