In both standard QuantSeq and QuantSeq-Pool library generation is initiated by oligo(dT) priming, where the primer contains a partial Illumina-compatible linker sequence. However, in QuantSeq-Pool this primer also contains Unique Molecular Identifiers (UMIs) and an i1 sample-barcode. This early barcoding allows the samples to be combined by pooling after first strand synthesis. All subsequent steps of the protocol can thus be performed on pools containing up to 96 samples. Batch processing significantly shortens the overall library generation time and reduces technical variability between samples within one pool.
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