Hello everyone, I am really new in Qiime2.
I have some stupid questions in importing data. Now I only have paired-end row data from illumina miseq. The format like this:
@MISEQ04:230:000000000-BLP2G:1:1101:12118:1920 1:N:0:TACATGTC
CTAGTAGGACCCTACGGGTGGCAGCAGTGGGGAATTTTTCGCAATTCTCTTTAGATTTACGCATCTACTCC
+
CCCCCGFAGGF@EGE777F:+C@,@86F,,C+,,CCEE,;+,,+8,<;@,,,,,,,<,,9,6,,6,,<,99,9,C,,,<,,6,,,,,99CF=,,,:C,C,,,,<C5
according to sequencing report, I realize in my raw data there exist single barcode(e.g. CTATAC) and primer sequence. I use mutiplexepairedEndBarcodeInsequence successfully imported my data and got an mutiplexed-seqs.qza. Then I want to use cutadapt to separate the samples sequences. Here is my code:
qiime cutadapt demux-paired \
--i-seqs multiplexed-paired-seqs.qza
--m-forward-barcodes-file metadata.tsv.xlsx
--m-forward-barcodes-column SequenceBarcode
--o-per-sample-sequences demultiplexed-seqs.qza
--o-untrimmed-sequences untrimmed.qza
but I got an error:
Error: Invalid value for "--i-seqs": File "multiplexed-paired-seqs.qza" does not exist.
I can't figure out the reason. What can I do ?