When I get a warning for base per sequence content, what should I do?

Authors: orcid logoFlorian Heyl avatar Florian HeylAdd Contributions!

So far it does not mean that your data is bad. Your protocol or your data might have a bias that you normally expect. Check first the following things:

  • Adapter content (maybe some adapters are still in your data)
  • Kmer content/Over represented sequences (this would indicate a contamination or a protocol/sequence bias)
  • Per base quality plot. If the overall quality is not good, then probably the sequencing was poorly performed.
  • Read about your protocol, e.g., ChIP-Seq and ATAC-Seq typically have a nucleotide bias. For example this article about ATAC-Seq.
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