Question: Should I select Workflow A or Workflow B for the i5 index sequence?
Answer: When sequencing the i5 index, Illumina instruments can use two different workflows:
- Workflow A = forward strand workflow
- Workflow B = reverse complement workflow
To determine whether your Illumina instrument uses the forward or reverse complement workflow, please refer to Illumina's Indexed Sequencing Overview Guide:
- The forward strand workflow is performed on the:
- NovaSeq 6000 with v1.0 reagent kits
- MiSeq
- MiniSeq with Rapid Reagent kits
- HiSeq 2500, HiSeq 2000
- The reverse complement workflow is performed on the:
- NovaSeq 6000 with v1.5 reagent kits
- iSeq 100
- MiniSeq with Standard reagent kits
- NextSeq Systems
- HiSeq X, HiSeq 4000, HiSeq 3000
In the CSV files for 10x Genomics Dual Index plates, both the Workflow A and Workflow B sequences are listed for the i5 sequence. Note that the Workflow B sequence is the reverse complement of the Workflow A sequence.
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