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  • Use same input amount

  • Compare samples at same read depth - down sample if necessary

  • Same Read 1 length

  • Do not collapse UMIs, even if UMIs are included in QuantSeq-FWD (with SSS UMI Module)

  • Recommend including technical replicates to assess handling variability (QSF samples processed separately whereas QSP samples processed together)

  • Recommend including controls (SIRVs) to assess DEG, if interested in this (compare expected vs observed)

What can also cause differences:

  • Differences in chemistries and protocol