09/22/21
Illumina offers a wide variety of library preparation kits and sequencing platforms for different applications, each with its own coverage and indexing considerations. The table below lists the sequencing primers included in the NovaSeq reagent cartridges, and the Illumina library preparation kits compatible with the NovaSeq platform based on the required data output and the indexing capabilities of each kit.
Note: The information in the tables below is based on the required data output and indexing capabilities of each kit. Most Illumina libraries can be sequenced with all Illumina sequencing platforms. If a specific Illumina library is not listed below, contact Illumina Tech Support to discuss special considerations.
Refer to the Sequence Coverage Calculator and Library Prep and Array Kit Selector for additional information.
| NovaSeq 6000 SP, S1, S2, and S4 v1 Reagent Kits | |||||
|---|---|---|---|---|---|
| Flow Cell Type |
Read 1 Primer |
Index 1 (i7) Primer |
Index 2 (i5) Primer |
Read 2 Primer |
Library Prep Kit |
| SP, S1, S2, S4 |
BP10 (Position 24 on cluster cartridge) |
BP14 (Position 23 on cluster cartridge) |
Grafted P5 on flowcell |
BP11 (Position 13 on cluster cartridge) |
Nextera DNA Exome |
|
Illumina DNA Prep (M) Tagmentation (formerly Nextera DNA Flex Library Prep Kit) |
|||||
|
Illumina DNA Prep with Enrichment (S) Tagmentation (formerly Nextera Flex for Enrichment) |
|||||
| Illumina Stranded mRNA, Ligation | |||||
|
Illumina Stranded Total RNA with Ribo-Zero Plus, Ligation |
|||||
|
Illumina RNA Prep with Enrichment (L) Tagmentation |
|||||
| TruSeq DNA Methylation | |||||
| TruSeq DNA PCR-Free | |||||
| TruSeq Exome | |||||
| TruSeq Nano DNA | |||||
| TruSeq RNA Access | |||||
| TruSeq Rapid Exome | |||||
| TruSeq Stranded Total RNA | |||||
| TruSeq Stranded mRNA | |||||
| TruSight Oncology 500 | |||||
| TruSight Oncology 500 ctDNA | |||||
| TruSight Oncology UMI | |||||
| SureCell WTA 3' | |||||
Table 1. Sequencing primer compatibility on the NovaSeq platform with NovaSeq v1 reagents, based on recommended library and flow cell types.
| NovaSeq 6000 SP, S1, S2, and S4 v1.5 Reagent Kits | |||||
|---|---|---|---|---|---|
| Flow Cell Type |
Read 1 Primer |
Index 1 (i7) Primer |
Index 2 (i5) Primer |
Read 2 Primer |
Library Prep Kit |
| SP, S1, S2, S4 |
VP10 (Position 24 on cluster cartridge) |
VP14 (Position 23 on cluster cartridge) |
BP11 (Position 13 on cluster cartridge) |
Nextera DNA Exome |
|
|
Illumina DNA Prep (M) Tagmentation (formerly Nextera DNA Flex Library Prep Kit) |
|||||
|
Illumina DNA Prep with Enrichment (S) Tagmentation (formerly Nextera Flex for Enrichment) |
|||||
| Illumina Stranded mRNA, Ligation | |||||
|
Illumina Stranded Total RNA with Ribo-Zero Plus, Ligation |
|||||
|
Illumina RNA Prep with Enrichment (L) Tagmentation |
|||||
| TruSeq DNA Methylation | |||||
| TruSeq DNA PCR-Free | |||||
| TruSeq Exome | |||||
| TruSeq Nano DNA | |||||
| TruSeq RNA Access | |||||
| TruSeq Rapid Exome | |||||
| TruSeq Stranded Total RNA | |||||
| TruSeq Stranded mRNA | |||||
| TruSight Oncology 500 | |||||
| TruSight Oncology 500 ctDNA | |||||
| TruSight Oncology UMI | |||||
| SureCell WTA 3' | |||||
Table 2. Sequencing primer compatibility on the NovaSeq platform with NovaSeq v1.5 reagents, based on recommended library and flow cell types.
For instructions on how to use custom primers on the NovaSeq sequencing platform, refer to the following resources: