Sequencing primer compatibility of Illumina libraries and recommended library types for the NextSeq 500/550 and MiniSeq sequencing platforms

09/20/21


Illumina offers a wide array of library preparation kits and sequencing platforms for different applications, each with its own coverage and indexing considerations. Table 1 and Table 2 list the sequencing primers included in the NextSeq 500/550 and MiniSeq reagent cartridges, and include the Illumina library preparation kits compatible with the NextSeq and MiniSeq platforms 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 resources for additional information.

Table 1. Sequencing primer compatibility with Illumina library prep kits recommended for the NextSeq 500/550 sequencing platform.

NextSeq 500/550

Read 1 Primer Index 1 (i7) Primer Index 2 (i5) Primer Read 2 Primer Library Type
BP10
(Position 20)
BP14
(Position 22)
BP11
(Position 21)
AmpliSeq for Illumina Library PLUS
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 RNA Prep with Enrichment (L) Tagmentation + Exome Panel
Illumina RNA Prep with Enrichment (L) Tagmentation + Respiratory Virus Oligo Panel
Illumina Stranded mRNA, Ligation
Illumina Stranded Total RNA with Ribo-Zero Plus, Ligation
Nextera XT
Respiratory Pathogen ID/AMR Enrichment Kit
TruSeq ChIP
TruSeq DNA Exome
TruSeq DNA Nano
TruSeq DNA PCR-Free
TruSeq Methyl Capture EPIC
TruSeq RNA Exome
TruSeq RNA v2
TruSeq Small RNA
TruSeq Stranded mRNA
TruSeq Stranded Total RNA
TruSeq Targeted RNA Expression
TruSight Cardio Sequencing Panel
TruSight Inherited Disease Sequencing Panel
TruSight Myeloid Sequencing Panel
TruSight Oncology 500
TruSight Oncology UMI
TruSight One Sequencing Panel
TruSight Rapid Capture Panels
TruSight RNA Fusion
TruSight RNA Pan-Cancer
TruSight Tumor 170

 

Table 2. Sequencing primer compatibility with Illumina library prep kits recommended for the MiniSeq sequencing platform.

MiniSeq
Read 1 Primer Index 1 (i7) Primer Index 2 (i5) Primer Read 2 Primer Library Type
BP10
(Position 24)
BP14
(Position 28)
BP11
(Position 25)
AmpliSeq for Illumina Library PLUS
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 RNA Prep with Enrichment (L) Tagmentation + Respiratory Virus Oligo Panel
Nextera XT
Respiratory Pathogen ID/AMR Enrichment Kit
TruSeq ChIP
TruSeq DNA Exome
TruSeq DNA Nano
TruSeq DNA PCR-Free
TruSeq Methyl Capture EPIC
TruSeq RNA Exome
TruSeq Small RNA
TruSeq Targeted RNA Expression
TruSight Cardio Sequencing Panel
TruSight Inherited Disease Sequencing Panel
TruSight Myeloid Sequencing Panel
TruSight One Sequencing Panel
TruSight Rapid Capture Panels
TruSight RNA Fusion
TruSight RNA Pan-Cancer
TruSight Tumor 15

 

For instructions to use custom primers on the NextSeq 500/550 and MiniSeq sequencing platforms, refer to the following resources: