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Fast QC per base sequence problem

i run Fast QC on sample using DNBSEQ and fail on per base sequence content but every remain result is okay

the result

And I think hexamer would not happen in this case so it is based on the enzyme cutting or smt else right ?

fastqc

Looks perfectly normal to me

those spiky lines in the beginning are very common and nothing to worry about (moreover: it is also a binning issue, first 9 bases is per base, after that they are 'grouped' per 5 , if you want you can disable this behaviour , --no_group or such?)

I would , personally, be more worried about all other metrics being 'green' ... that is not so common

Keep in mind that FastQC is a tool that has been around for quite a while and is actually not designed to accurately work with the modern day sequencing output(s)

And I think hexamer would not happen in this case so it is based on the enzyme cutting or smt else right ?

Positional/sequence bias is likely present (in spite of kits claiming its absence) and that is what you are likely seeing at the beginning of reads. Sounds like you have already read this blog --> https://sequencing.qcfail.com/articles/positional-sequence-bias-in-random-primed-libraries/ .

Plot is suggesting a AT rich library of some sort. What kind of sequencing is this? Perhaps your organism has AT rich transcriptome, this is a poly-A enriched library or a total RNAseq library (rRNA present in large amounts).

Very tks for u guys that have answered me, i would provided some information if u need it: it is a read of DNA of black soldier fly SRR34006749

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