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| Product Name | Next-Generation Sequencing (NGS) Adapter Oligonucleotide Set |
| Catalog No. | PRIPRO-0022 |
| Description | A custom-synthesized set of oligonucleotides for constructing next-generation sequencing (NGS) library adapters. NGS adapters contain platform-specific sequences required for cluster generation or bead loading, library amplification, and index (barcode) sequences for sample multiplexing. This service synthesizes adapter oligonucleotides to customer-specified sequences, including the P5/P7 flow cell binding sequences for Illumina platforms, Ion Torrent adapter sequences (A and P1), or custom adapter designs for other platforms. Adapters may include single-index or dual-index (unique dual indexing, UDI) configurations. Typical adapter design includes a double-stranded region for ligation via T/A overhang with a Y-shaped or stem-loop structure to enable directional sequencing. All oligos are HPLC or PAGE purified and supplied as matched pairs ready for annealing. |
| Intended Use | Designed for NGS library preparation across Illumina (NovaSeq, HiSeq, MiSeq, NextSeq), Ion Torrent (PGM, Proton, S5), and other sequencing platforms; suitable for whole-genome, exome, RNA-seq, ChIP-seq, ATAC-seq, amplicon-seq, and custom targeted sequencing applications. |
| Principle / Technology | Custom oligonucleotide synthesis of platform-specific adapter sequences; adapters ligated to end-repaired, A-tailed DNA fragments; indexed adapters enable sample multiplexing; Y-shaped structure enables directional sequencing and prevents adapter dimer formation |
| Detection Method | HPLC or PAGE purification; MALDI-TOF MS verification; functional QC by test ligation and PCR amplification; annealing of matched pairs verified by non-denaturing PAGE |
| Sample Type | End-repaired and A-tailed DNA fragments from genomic DNA, cDNA, or PCR amplicons; applicable to a wide range of input amounts (1 ng – 1 µg) |
| Performance Range / Specifications | Custom adapter sequences 30–70 bases; 5′ phosphorylation available; phosphorothioate bonds for nuclease resistance; dual-index configurations with 6–12 base barcodes |
| Sensitivity / LOD | Enables library preparation from as low as 50 pg input DNA with optimized protocol; adapter dimer contamination <0.1% by Bioanalyzer when properly HPLC purified |
| Specificity | Sequence-perfect adapter oligos by MS verification; matched pairs anneal with >98% efficiency; index sequences with minimum Hamming distance ≥3 for error-tolerant demultiplexing |
| Reaction Conditions / Protocol | Anneal matched adapter oligos at equimolar concentration (10–50 µM each) in annealing buffer (10 mM Tris pH 8.0, 50 mM NaCl, 1 mM EDTA): heat to 95 °C for 5 min, cool gradually to 25 °C over 45–60 min. Use annealed adapter at 1.5–15 µM in ligation reaction. |
| Components / Formulation | Matched pair of adapter oligonucleotides (e.g., P5-index + P7-index or A + P1); supplied as separate tubes for custom annealing or pre-annealed duplex upon request; COA for each oligo |
| Storage Conditions | –20 °C for individual oligos; –20 °C for annealed adapter aliquots |
| Shelf Life | 24 months at –20 °C dry oligos; 12 months annealed at –20 °C |
| Package Specifications | Two or more tubes per adapter set; 10–100 nmol scale per oligo; custom scales available; pre-annealed option in single tube |
| Product Form | Lyophilized white pellets (individual oligos) or clear solution (pre-annealed duplex) |
| Quality Control | HPLC or PAGE purity ≥90% per oligo; MALDI-TOF MS mass confirmation; functional test ligation and library amplification generating expected Bioanalyzer trace; residual adapters and dimers <1%; endotoxin testing available. |
| Key Features | Custom adapter design and synthesis; platform-specific sequences (Illumina, Ion Torrent, or custom); single or dual indexing; HPLC purified for minimal dimerization; Y-shaped design for directional sequencing; pre-annealed option available; design consultation provided. |
| Purity | ≥90% HPLC or ≥95% PAGE per individual oligo; <1% adapter dimer in annealed product |
| Concentration | Individual oligos: resuspend to 100 µM stock; annealed adapter: supplied at 15 µM or 50 µM as duplex; A260 verified |
| Activity / Unit Definition | Ligation efficiency >90% (molecules ligated per input); PCR amplification of adapter-ligated fragments successful with 6–15 cycles; minimal adapter dimer amplification |
| Molecular Weight | Sequence-dependent; each oligo mass reported on COA; verified by MS |
| Source / Origin | Synthetic phosphoramidite chemistry; no biological templates |
| pH Range / Optimal pH | Stable pH 6.0–9.0; annealing buffer pH 7.5–8.0 optimal |
| Shipping Conditions | Ambient temperature as lyophilized oligos; cold pack for pre-annealed solution; express delivery available |
| Expiration Date / Stability | 24 months dry at –20 °C; 12 months annealed at –20 °C; aliquot annealed adapters to avoid repeated freeze-thaw |
| Regulatory / Compliance | For research use only; manufactured under ISO 9001; not for clinical diagnostic sequencing without validation |
| Compatibility | Compatible with commercial library preparation kits from major suppliers (NEBNext, KAPA HyperPrep, TruSeq-compatible, Ion Xpress); compatible with all vendor sequencing instruments |
| Recommended Buffer System | Annealing buffer: 10 mM Tris-HCl pH 8.0, 50 mM NaCl, 1 mM EDTA; compatible with T4 DNA ligase buffer for ligation reactions |
| Application Notes / Precautions | Annealed adapters should be aliquoted and stored at –20 °C. Molar ratio of adapter to insert is critical for efficient ligation — follow kit recommendations (typically 10:1 to 100:1). For low-input libraries, reduce adapter concentration to minimize dimer formation. |
| Batch-to-Batch Consistency | Adapter ligation efficiency within ±10% across batches; MS mass accuracy ±0.05%; Bioanalyzer library profile consistent across production lots |
For research use only, not for clinical use.
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