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Single-read 50 (SR50) sequencing runs are suitable for both QuantSeq versions. Although, for QuantSeq REV it is favorable to use longer runs (SR100), since starting the read exactly at the 3`UTR will also show the polyadenylation signal, which covers up to 25 nt from the transcription end. As the polyadenylation signal is quite similar for all transcripts, it is better to have longer reads >SR50 available for mapping in order to increase the number of uniquely mapping reads.