In both LUTHOR HD and LUTHOR HD Pool, library generation is initiated by oligo(dT) priming, where the primer contains a 12 nt Unique Molecular Identifier (UMI) and a partial Illumina-compatible linker sequence. However, in LUTHOR HD Pool, this primer also contains a 12 nt i1 sample-barcode.
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For best practice, we would recommend keeping kit components separate and avoid mixing/matching. Importantly, IVT and E4 are NOT interchangeable between kits. Additionally, some steps between protocols may differ e.g., LUTHOR HD Pool includes an additional incubation step during purification THOR reaction (Step 7; bold): 10 minutes at 37 °C, 5 minutes at 42 °C, 20 minutes at 65 °C, then cool to 25 °C and hold for 1 minute.