Indexing FAQ for Illumina DNA PCR-Free libraries

If there is no PCR, how are the indexes and full adapter sequences incorporated during library preparation? There is a proprietary mechanism that allows for ligation of Illumina DNA/RNA Unique Dual (UD) index primers.

Can Unique Molecular Identifiers (UMIs) be incorporated into this workflow? No.

Can UMI anchors be used with the Illumina DNA PCR-Free library prep kit? No, they are incompatible.

As the protocol uses only half of the volume in the adapter plate (5 µl out of an approximately 10 µl fill volume), why shouldn't the index plate wells be reused? This is to prevent cross-contamination and ensure that there is sufficient volume available for use.

**Can custom index adapters or primers be used with the Illumina DNA PCR-Free kit?**No.

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