Sequencing primer compatibility of Illumina libraries and recommended library types for the MiSeq sequencing platform

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 lists the sequencing primers included in the MiSeq reagent cartridges, and the Illumina library preparation kits compatible with the MiSeq platform.

Note: The information in the table 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 primers for the recommended library types for the MiSeq sequencing platform.

MiSeq

Read 1
Primer

Index 1
(i7)
Primer

Index 2
(i5)
Primer

Read 2
Primer

Library Type

HP10
(position 12)

HP12
(position 13)

Grafted P5
on flow cell

HP11
(position 14)

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 Nano
TruSeq DNA PCR-Free
TruSeq RNA v2
TruSeq Small RNA
TruSeq Stranded mRNA
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 on how to use custom primers on the MiSeq sequencing platform, refer to the following resources: