Indexing FAQ for Illumina DNA PCR Free libraries
If there is no PCR, how indexes and full adapter sequences incorporated during library preparation? There is a proprietary mechanism in Illumina DNA PCR-Free allows for ligation of IDT for Illumina - DNA/RNA Unique Dual (UD) index primers.
Can Unique Molecular Identifier (UMIs) be incorporated into this workflow? This has not been validated.
Can UMI anchors (IDT for Illumina - UMI DNA/RNA UD Indexes, Ligation (formerly known as IDT for Illumina - UMI DNA Index Anchors)) be used with the Illumina DNA PCR-Free library prep kit? No, the UMI anchors/index sets cannot be used with Illumina DNA PCR-Free Prep, Tagmentation library prep kit.
As the protocol uses half the volume in the adapter plate (5 µl of approximate 10 µl fill), why does the protocol say the index plate cannot be reused? This is to prevent cross-contamination and make sure that there is sufficient volume for use.
Can custom index adapters with the Illumina PCR-Free kit?
No.
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