- NNEXTFLEX® Dual-Indexed Adapter (25 µM)
- NEXTFLEX® Primer Mix (12.5 µM)
- Sequences of NEXTflex Dual-Indexed DNA Barcode Indexes – Excel / PDF
The NEXTFLEX® Dual-Indexed DNA Barcodes contain 8 reactions each of 96 unique adapters compatible with Illumina® platforms, enabling the user to multiplex up to 192 samples. This kit is shipped on dry ice.
What Sequencing Primers Do I Use for Dual-Indexed Libraries?
Libraries made with the NEXTFLEX® Dual-Indexed DNA Barcodes differ from libraries made with the NEXTFLEX® DNA Barcodes in that they have two index sequences, as opposed to a single barcode index. Depending on the sequencing platform and flowcell being used, dual-indexed libraries may require the use of special sequencing primers in place of the primers found in the TruSeq® Cluster Kit.
Sequencing of dual-indexed libraries on the MiSeq® does not require the use of the TruSeq® Dual Index Sequencing Primer Box in addition to the MiSeq® Reagent Kit.
Sequencing dual-indexed libraries on the HiSeq® or GAIIx, however, may require the use of the TruSeq® Dual Index Sequencing Box. The TruSeq® Dual Index Sequencing Box, SR (Illumina® cat. # FC-121-1003) is required when sequencing dual-indexed libraries on a single-read flow cell. In this case, the Index 2 (i5) Sequencing Primer Mix, HP9, must be loaded in position 16 on the flowcell. If sequencing dual-indexed libraries on a paired-end flow cell for HiSeq® or GAIIx, the TruSeq® Dual Index Sequencing Box is not required, as the Index 2 primer is grafted on the flow cell.
In summary, sequencing dual-indexed libraries requires additional primers only if sequencing on a single-read flow cell on the HiSeq® or GAIIx. Sequencing on a paired-end flow cell on the HiSeq® or GAIIx or on the MiSeq® does not require additional primers to be purchased in addition to the typical sequencing primer kit for each sequencer.