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 HiSeq X and HiSeq 3000/4000 reagent kits, and include the Illumina library preparation kits compatible with the HiSeq X and HiSeq 3000/4000 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 HiSeq X sequencing platform.
HiSeq X |
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Read 1 |
Index 1 |
Index 2 (i5) |
Read 2 |
Library Type |
HP10 |
HP12 |
N/A |
HP11 |
Illumina DNA Prep (M) Tagmentation (formerly Nextera DNA Flex Library Prep Kit, 350 bp insert size only) TruSeq DNA Nano (350 bp insert size only) TruSeq DNA PCR-Free (350 bp insert size only) |
Table 2. Sequencing primer compatibility with Illumina library prep kits recommended for the HiSeq 3000/4000 sequencing platform.
HiSeq 3000/4000 |
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Flow cell Type |
Read 1 |
Index 1 |
Index 2 |
Read 2 |
Library Type |
Single Read |
HP10 |
HP12 |
Grafted P5 on |
N/A |
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) Nextera XT TruSeq ChIP TruSeq DNA Exome TruSeq DNA Nano (350 bp insert size only) TruSeq DNA PCR-Free (350 bp insert size only) TruSeq Methyl Capture EPIC TruSeq RNA Exome TruSeq RNA v2 TruSeq Stranded mRNA TruSeq Stranded Total RNA TruSight Oncology UMI TruSight One Sequencing Panel |
Paired End |
HP10 |
HP14 |
HP11 |
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For instructions to use custom primers on the HiSeq 3000/4000 sequencing platforms, refer to the following resources: