My data is sequenced using Illumina NovaSeq 6000 and another dataset using Illumina MiSeq so running dada2 wasn't successful so I tried to use deblur instead. Before getting to the error faced while running deblur, is there a way to use dda2 on such data?
Both showed the same error using the deblur tool but different errors using dada2.
When I tried to run this command for one of them:
qiime deblur denoise-16S \
--i-demultiplexed-seqs ch1-demux-filtered.qza
--p-trim-length 249
--o-representative-sequences ch1-rep-seqs-deblur.qza
--o-table ch1-table-deblur.qza
--p-sample-stats
--o-stats ch1-deblur-stats.qza
although the files are there in the same directory, including the files from the filtering step
When I tried to run the same data through dada2 this what I faced for the two of them:
This is the paried-end dataset using Illumina NovaSeq 6000 (but only the forward reads is being used)
As for the second DADA2 error, the importing step was done successfully with no errors reported, and the data is already published so I don't think it can be corrupted?
I am attaching the demux file for your reference. Your help is greatly appreciated. single-end-demux.qzv (292.5 KB)
Thank you for the clarification. However, I just want to highlight that the two errors are for two different datasets. I have read somewhere on the forum that DADA2 can't work with reads sequenced by higher sequencers such as Illumina NovaSeq 6000 and Illumina MiSeq (which is my case). The dataset with the first DADA2 error worked fine with deblur but the second showed an error which I am still trying to fix. Does the dataset's successful work with deblur mean that it's not corrupted? Or this has nothing to do with that?
At this point I am totally lost, the demux report you'd send doesn't look like raw data. If it's downloaded from SRA there are many things that could go wrong. I personally have pointed out a few to SRA maintainers, but the database is too big to fix everything.
It might be that sequences were just quality controlled before, but I don't know.
I had processed sequences from MiSeq before in DADA2 and had no problems with it.
deblur does not rely on quality scores but only on the sequences, that is why it can be used to co-analyze samples from different runs as well as samples from NovaSeq platform (which does bin the quality scores). So you should be able to process all the samples together with deblur.
Sorry, I lost the track of the error now, what is your latest error you are working on? Lets see if we can help more.
Cheers and happy new year
Luca