
High quality adapters that meet the ever-expanding needs of researchers using NGS
Adapters are a key component of the next generation sequencing (NGS) workflow. Whether your project requires a basic adapter or a sophisticated design for higher accuracy, IDT has the products and expertise to deliver the right solution.
Regardless of the NGS instrument or application, IDT has been serving the needs of NGS scientists for over 10 years. We are recognized as the leader in adapter synthesis based on our experience in custom oligo manufacturing and our commitment to quality. Additionally, our NGS customers and partners benefit from:
Due to our leadership in NGS adapter synthesis, Illumina chose IDT as its partner to develop the next generation of index adapters to improve sample multiplexing. These adapters contain Illumina’s unique dual indexes (UDIs) that mitigate sample misassignment due to index hopping. IDT manufactures UDI adapters for Illumina and is also licensed to sell your own custom designed adapters containing these UDI sequences. See the press release here.
Figure 1. Examples of adapter designs. A variety of NGS adapter designs are available. When selecting or designing adapters, consideration must be given to the intended application, multiplexing needs, accuracy requirements, and analysis methods.
IDT provides xGen Dual Index UMI Adapters, xGen Stubby Adapter and UDI Primer Pairs, and a Custom Adapter Configurator tool that guides you through design of Custom NGS Adapters. Below we describe some of the design considerations you will want to take into account.
Sequences for specific NGS platforms: During library preparation, adapters are attached (by ligation, PCR, or tagmentation) to the fragments of each sample library. Adapters include platform-specific sequences for fragment recognition by the sequencer: for example, the P5 and P7 sequences (Figure 1) enable library fragments to bind to the flow cells of Illumina platforms. Each NGS instrument provider uses a specific set of sequences for this purpose. IDT manufactures adapters for all major NGS platforms.
Sample indexing: Sample indexes (or indices) enable multiple samples to be sequenced together (i.e., multiplexed) on the same instrument flow cell or chip. Each sample index, typically 6–10 bases, is specific to a given sample library and is used for de-multiplexing during data analysis to assign individual sequence reads to the correct sample. Adapters may contain single or dual sample indexes depending on the number of libraries combined and the level of accuracy desired. Illumina recommends using unique dual indexes (UDIs) as a method to mitigate errors introduced by index-hopping. UDIs are particularly important when using instruments with patterned flow cells, such as the NovaSeq system.
IDT provides several indexing options with its Custom NGS Adapters. These include both IDT and Illumina 8 and 10 base index series, as well as the ability to upload your own index files.
Molecular barcoding: Unique molecular identifiers (UMIs) provide the highest levels of error correction and accuracy. UMIs are short sequences, often with degenerate bases, that incorporate a unique barcode onto each molecule within a given sample library. UMIs have been shown to reduce the rate of false-positive variant calls and increase sensitivity of variant detection. By incorporating individual barcodes on each original DNA fragment, variant alleles present in the original sample (true variants) can be distinguished from errors introduced during library preparation, target enrichment, or sequencing. Any identified errors can be removed by bioinformatics methods before final data analysis. Adapters that contain UMIs, such as the xGen Dual Index UMI Adapters, are available with a UDI design for detection of low-frequency variants. IDT Custom NGS Adapters can also be configured with UMIs.
Adapter manufacturing: Stringent manufacturing methods are critical for producing high quality NGS adapters. Substandard manufacturing practices can lead to low purity adapters or adapter cross contamination, either of which will negatively impact sequencing results. The IDT proprietary TruGrade process uses state-of-the-art synthesis and purification methods, designed specifically for NGS adapters. IDT also offers GMP grade adapter manufacturing for clinical applications.
Choose IDT adapter products based on the flexibility your experimental workflow requires. Click on the xGen Dual Index UMI Adapters—Tech Access, the xGen Stubby Adapter and UDI Primer Pairs, or the Custom NGS Adapters headings in Table 1 to review product offerings. The Custom NGS Adapters provide a step-by-step tool to help you configure adapters best suited for your research.
Specifications | xGen Dual Index UMI Adapters—Tech Access | xGen Stubby Adapter and UDI Primer Pairs | Custom NGS adapters |
---|---|---|---|
Configuration | UDI-UMI | UDI | single index, combinatorial dual indexing, UDI, or UDI-UMI |
Sample index length | 8 base, non-redundant sample indexes | 8 base, non-redundant sample indexes | Configurable |
Concentration | 15 µM | 15 µM (Stubby Adapter), 20 µM total (10 µM each Primer) | 15 µM (Adapter), 20 µM total (10 µM each Primer) |
Shipping container | Tubes, plates | Tubes (Stubby Adapter), 96-well plates (UDI Primer Pairs) | Tubes, plates, or single-use plates |
Reactions | Varies | 1 reaction per primer pair | Varies |
Adapter modifications | Available | Methylated for bisulfite sequencing* | Available |
Illumina platform compatibility | 2- and 4-color platforms | 2- and 4-color platforms | Designs for any application |
* For bisulfite sequencing, see our FAQ for additional information.
One kit. Flexible workflow. Endless applications.
The Lotus DNA Library Prep Kit enables streamlined preparation of high-quality next generation sequencing (NGS) libraries from double-stranded DNA (dsDNA). The kit uses enzymatic fragmentation to generate libraries suitable for PCR-free, PCR-amplified, and targeted sequencing applications on Illumina platforms.
The Lotus kit can be customized for your applications when combined with one of the many IDT adapter options (see Ordering section). Additionally, use of xGen hybridization capture products provides a complete NGS solution that takes you from sample preparation to sequencing.
The Lotus DNA Library Prep Kit produces high-quality next generation sequencing (NGS) libraries that are suitable for many applications including:
This kit involves minimal enzymatic incubations and bead-based purification steps, thereby reducing sample handling and overall library preparation time to under 2 hours. The major steps are depicted in Figure 1 and are summarized as follows:
Figure 1. Overview of Lotus DNA library preparation. Our easy enzymatic method takes you from sample to sequencing while eliminating the need for acoustic shearing methods that require instrumentation and extra time.
The Lotus kit comes with all the buffers, reagents, and enzymes needed for fragmentation, adapter ligation, and an optional PCR (to add index sequences, depending on adapter type, or to increase the amount of DNA for sequencing). You supply the DNA and add adapters that fit your goals. Table 1 provides additional specifications for using this kit.
Table 1. Specifications of the Lotus DNA Library Prep Kit.
Feature | Details |
---|---|
Kit sizes | 16 reactions 96 reactions |
Sample types |
Double-stranded DNA from fresh, frozen tissue Genomic DNA Full-length, double-stranded cDNA |
DNA input range | 1–250 ng |
Suggested DNA insert size | Whole-genome applications: 350 bp Hybridization capture: 200 bp |
TA-ligation adapters (see Table 2) |
Full-length: PCR amplification is optional Stubby (truncated): PCR is required to add index sequences |
Complete your library prep with our selection of ready-to-use xGen Dual Index UMI Adapters or xGen Stubby Adapter and UDI Primer Pairs (Table 2). We also offer many high-quality, custom adapters and index primers that are application-specific and ideal for use with the Lotus kit.
Table 2. How to choose IDT adapters for common applications when using the Lotus DNA Library Prep Kit.
Application | Best choice | Rationale |
---|---|---|
Whole genome sequencing (WGS), metagenomics, PCR-free sequencing (≥100 ng input) | xGen Dual Index UMI Adapters–Tech Access | Full-length adapters – required for PCR-free applications |
WGS, metagenomics with PCR (1–250 ng input), exome sequencing, and targeted germline sequencing (SNVs, indels) | xGen Stubby Adapter and UDI Primer Pairs | Simple workflow – prepare your ligation master mix with a universal adapter and introduce unique dual index sample barcodes via PCR |
Low-level mutation detection, down to ~1% frequency | xGen Dual Index UMI Adapters–Tech Access | More sensitive – use UMI consensus analysis to remove sequencing errors |
RNA-seq starting with full-length, double-stranded cDNA input | xGen Dual Index UMI Adapters–Tech Access | More quantitative – UMI is used to remove PCR duplicates |
Lotus DNA Library Prep Kit paired with adapters from IDT create libraries that are compatible with xGen hybridization capture probes and reagents for when you only need to perform targeted sequencing (Figure 2). Here is a list of the key components you will need:
Go ahead and commit. We made a lot.
We designed this updated panel using our new target-aware algorithm, resulting in the most complete coverage of any exome panel on the market. We assessed each probe with proprietary off-target analysis and manufactured them under strict ISO 13485 standards. We performed electrospray ionization mass spectrometry (ESI-MS) and dual quantification on each of the 415,115 probes before pooling. Finally, individual probe synthesis means we can deliver on a much larger scale than array methods, and produce a single large lot that can be aliquoted and stored securely, for years of reproducibility.
The xGen Exome Research Panel v2 gives you the confidence that only individual probe synthesis and individual QC can provide, and it comes at a price that makes your choice easy.
Exome sequencing sequences only the protein-coding regions of the genome. Since exons represent only 1% of the human genome, an effective method of separating these regions from noncoding DNA is critical to focus on potentially important mutations implicated in disease. The captured material must also be suitable for sequencing to a satisfactory depth of coverage for reliable detection of variant alleles and sensitive applications like copy number variation (CNV) detection. To provide increased depth of coverage and enable high multiplexing of samples, the
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