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Cat. No.: BADIPT-1 (for Set 01 (96 reactions, 24 index pairs))

Cat. No.: BADIPT-2 (for Set 02 (96 reactions, 24 index pairs))

Cat. No.: BADIPT-3 (for Set 03 (96 reactions, 24 index pairs))

Cat. No.: BADIPT-4 (for Set 04 (96 reactions, 24 index pairs))

Description

SBS-NGS™ Bidirect Amp Dual-Index Primers (For Illumina) is a specialized dual-index PCR primer kit developed for amplicon sequencing (AmpliSeq) library construction on the Illumina sequencing platform. It is designed for use with our series of SBS-NGS™ amplicon sequencing kits, such as the SBS-NGS™ Human Mitochondrial DNA Mutation Detection Kit (For Illumina). With this kit, users can reliably and efficiently generate high-throughput sequencing libraries compatible with Illumina platforms.

The SBS-NGS™ Bidirect Amp Dual-Index Primers (For Illumina) kit includes four sets (Set01–Set04). Each set contains four 8-base (nt) i5-tagged P5 primers and six 10-base (nt) i7-tagged P7 primers. When used individually, each set allows cross-combination of P5 and P7 primers to generate 24 unique i5/i7 index pairs. When all four sets are used together, up to 384 unique i5/i7 index pairs can be generated, greatly expanding the multiplexing capacity of amplicon sequencing libraries.

Each set (Set01–Set04) of the SBS-NGS™ Bidirect Amp Dual-Index Primers (For Illumina) kit contains two primer groups (Set A and Set B):

• Set A: Forward Strand PCR Dual-Index Primers, including 4 upstream primers (SBS-NGS™ Amp P5 Primer A) and 6 downstream primers (SBS-NGS™ Amp P7 Primer A).

• Set B: Reverse Strand PCR Dual-Index Primers, including 4 upstream primers (SBS-NGS™ Amp P5 Primer B) and 6 downstream primers (SBS-NGS™ Amp P7 Primer B).

Within the same series, Set A and Set B use the same 24 i5/i7 index pairs; however, primers from Set A and Set B must not be mixed. The detailed PCR amplification principle and usage strategy are illustrated in Figure 1, and the i5/i7 index sequence information can be found in FAQ 1 of this manual.

Figure 1. Amplification strategy and schematic of the SBS-NGS™ Bidirect Amp Dual-Index Primers (For Illumina).

Sequencing libraries constructed using the SBS-NGS™ Bidirect Amp Dual-Index Primers (For Illumina) kit can be sequenced directly on Illumina platforms without the need for additional sequencing primers.

Performance testing has demonstrated that the SBS-NGS™ Bidirect Amp Dual-Index Primers (For Illumina) kit delivers highly uniform sequencing output. For the same sample amplified with different index primer pairs, repeated sequencing runs showed less than 5% variation in data yield. To ensure optimal uniformity across sequencing samples, users are advised to maintain high DNA sample quality and minimize amplification efficiency differences during the initial target region amplification step.

Each set of the SBS-NGS™ Bidirect Amp Dual-Index Primers (For Illumina) kit supports 96 library indexing amplification reactions.

Components

Set 01

• SBS-NGS™ Nex i5 Primer 01
• SBS-NGS™ Nex i5 Primer 02
• SBS-NGS™ Nex i5 Primer 03
• SBS-NGS™ Nex i5 Primer 04
• SBS-NGS™ Nex i7 Primer 01
• SBS-NGS™ Nex i7 Primer 02
• SBS-NGS™ Nex i7 Primer 03
• SBS-NGS™ Nex i7 Primer 04
• SBS-NGS™ Nex i7 Primer 05
• SBS-NGS™ Nex i7 Primer 06

Set 02

• SBS-NGS™ Nex i5 Primer 05
• SBS-NGS™ Nex i5 Primer 06
• SBS-NGS™ Nex i5 Primer 07
• SBS-NGS™ Nex i5 Primer 08
• SBS-NGS™ Nex i7 Primer 07
• SBS-NGS™ Nex i7 Primer 08
• SBS-NGS™ Nex i7 Primer 09
• SBS-NGS™ Nex i7 Primer 10
• SBS-NGS™ Nex i7 Primer 11
• SBS-NGS™ Nex i7 Primer 12

Set 03

• SBS-NGS™ Nex i5 Primer 09
• SBS-NGS™ Nex i5 Primer 10
• SBS-NGS™ Nex i5 Primer 11
• SBS-NGS™ Nex i5 Primer 12
• SBS-NGS™ Nex i7 Primer 13
• SBS-NGS™ Nex i7 Primer 14
• SBS-NGS™ Nex i7 Primer 15
• SBS-NGS™ Nex i7 Primer 16
• SBS-NGS™ Nex i7 Primer 17
• SBS-NGS™ Nex i7 Primer 18

Set 04

• SBS-NGS™ Nex i5 Primer 13
• SBS-NGS™ Nex i5 Primer 14
• SBS-NGS™ Nex i5 Primer 15
• SBS-NGS™ Nex i5 Primer 16
• SBS-NGS™ Nex i7 Primer 19
• SBS-NGS™ Nex i7 Primer 20
• SBS-NGS™ Nex i7 Primer 21
• SBS-NGS™ Nex i7 Primer 22
• SBS-NGS™ Nex i7 Primer 23
• SBS-NGS™ Nex i7 Primer 24

Storage

Store at –20 °C. Stable for at least one year.

Notes

• Required reagents and materials (not included): DNA size-selection magnetic beads, Ultrapure Water (DNase/RNase-Free, Sterile), magnetic separation rack, 96-/384-well PCR instrument, 96- or 384-well PCR plates, and heat-resistant PCR sealing film. These can also be purchased from us if needed.
• All primers in this product are single-stranded oligonucleotides. Do not expose them to temperatures above room temperature, and avoid repeated freeze–thaw cycles to ensure optimal performance.
• Follow best laboratory practices to prevent cross-contamination of indexed adapters.
• This product is intended for research use only by trained professionals. It must not be used for clinical diagnosis or therapy, food, or pharmaceuticals, and should not be stored in residential settings.
• For your safety and health, always wear a lab coat and disposable gloves when handling this product.

Related:

SBS-NGS™ Bidirect Amp Dual-Index Primers (For Illumina)