Weird 8bp peak in (clean) small RNA data
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2.5 years ago

Hi everyone,

I've received some small RNA sequencing data from a collaborator and after removing adapter and trimming (with TrimGalore) i have a small peaks at ~20 and 30 bp, as I was expecting, plus a huge peak at 8bp that contains most of the bases of the sequencing. There are no overepresented sequences in the samples and they used the NEXTFLEX® Small RNA-seq. This is a whole insect sample. I saw a lot of unexpected lenght peaks in small RNA sequencing but never at 8bp and I have no idea what that is, it is present in all the samples they received. Does anyone with more experience than me know what this 8bp sequences may be? Thanks and have a nice day!!

fastqc graph for the read length

NGS small-RNA • 597 views
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It could be just be "bad" data... and the majority of their RNA was degraded or something leading to an enrichment of 8-nt RNA fragements... (I'm assuming these are trimmed so that means the rest of the read was barcode).

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What did the sequence length distribution look like for the raw data?

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