Twist Bioscience NGS | Library Preparation | Universal Adapter Systems
Twist Bioscience Next Generation Sequencing Library Preparation
Universal Adapter Systems
Twist’s Universal Adapter Systems improve the performance of any ‘T-A’ overhang ligation library preparation workflow for next-generation sequencing (NGS) applications. Engineered to optimize the performance of Twist’s Library Preparation and Target Enrichment products, the systems consist of Twist’s Universal Adapters and either Twist Unique Dual-Indexed (UDI) Primers or Twist HT UDI Primers.
Twist Universal Adapters
The proprietary design of the universal TruSeq-compatible adapters ensures compatibility with all ‘T-A’ overhang workflows and eliminates formation of undesired adapter dimer. It also contributes to increasing the workflow’s overall yield, which is especially beneficial prior to a target enrichment step.
Twist 10 bp Unique Dual-Indexed (UDI) Primers
Twist’s UDI primers contain mixed primer pairs, which enable higher confidence sample pooling than combinatorial approaches. The index design allows for correction of substitutions, insertions, and deletions within the index reads during post-sequencing processing.
Twist 12bp High Throughput (HT) UDI Primers
Available in automation-friendly 96-well or 384-well plate formats, Twist’s HT UDI primers offer the most scalable solution for high throughput sequencing. All 3,072 unique indexes take into consideration base and color channel balancing, making them optimal for either two-color or four-color chemistries.
- Drastically reduce adapter dimer formation
- Use fewer PCR cycles during library preparation to reduce errors and bias introduced during amplification
- Can be used with all ‘T-A’ overhang ligation systems
- Compatible with all Illumina sequencing chemistries (2- or 4-color) and flow cell types (non-patterned and patterned)
- Scale your multiplexing with up to 3,072 available indexes
- Ready-to-use formulation
- Reduces per-sample sequencing cost
Increase Final Yields During Library Preparation
Twist Universal Adapters and HT Universal Adapters return higher total yields with fewer PCR cycles. This ultimately leads to the production of more library with fewer PCR errors to drive improvements in target enrichment efficiency.”
The chart to the right shows how using Twist Universal Adapters for insert ligation and UDI primer PCR amplification generates a robust library.
Library preparation and target enrichment work together to influence the ability to call variants in sequencing. Consistently constructing robust libraries is important for successful hybrid capture. When adapters dimerize during library preparation, they generate undesired products and reduce the effective concentration of adapters available for ligation to inserts. Using Twist Universal Adapters and Primers during library construction minimizes this problem.
Twist Universal Adapters minimize adapter dimer formation during library preparation.
DOWNLOADS
The Twist Universal Adapter Systems improve the overall performance of any ‘T-A’ overhang ligation library preparation workflow for next-generation sequencing (NGS) applications. Read the product sheet to learn more.
- Key benefits of the Twist Universal Adapter System
- How the Universal Adapter Systems design leads to optimal performance
PROTOCOLS
EF 1.0 Library Preparation and Universal Adapter System
This protocol is for the preparation of genomic DNA (gDNA) libraries using enzymatic gDNA fragmentation with either the Twist Universal Adapter System or the Twist HT (High-Throughput) Universal Adapter System. The Twist Universal Adapter System and Twist HT Universal Adapter System improve the overall performance of any ‘T-A’ overhang ligation library preparation workflow for next-generation sequencing (NGS) applications. This manual details the steps for generating the amplified, indexed libraries needed for downstream target enrichment and sequencing on Illumina NGS systems. This library preparation protocol is optimized for use with Twist Target Enrichment Kits and should only be performed with reagents specified or their equivalents.
- User Supplied Materials and Equipment
- General Notes and Precaution
- Guidelines for gDNA Sample
- Twist Enzymatic Fragmentation Library Preparation Workflow
This Twist Library Preparation Kit provides the reagents needed to prepare genomic DNA (gDNA) libraries using enzymatic gDNA fragmentation with either the Twist Universal Adapter System or the Twist HT (High-Throughput) Universal Adapter System. The Twist Universal Adapter System consists of Twist Universal Adapters and Twist Unique Dual Indexed (UDI) Primers, whereas the Twist HT Universal Adapter System includes Twist Universal Adapters and HT UDI Primers. This manual details the steps for generating the amplified, indexed libraries needed for downstream target enrichment and sequencing on Illumina next-generation sequencing (NGS) systems. This library preparation protocol is optimized for use with Twist Target Enrichment Kits and should only be performed with reagents specified or their equivalents.
- User Supplied Materials and Equipment
- General Notes and Precaution
- Guidelines for gDNA Sample
- Twist Enzymatic Fragmentation 2.0 Library Preparation Workflow
This protocol is for the generation of amplified, indexed libraries needed for downstream target enrichment and sequencing on Illumina next-generation sequencing (NGS) systems. This Twist Library Preparation Kit provides the reagents needed to prepare genomic DNA (gDNA) libraries using mechanically fragmented gDNA with either the Twist Universal Adapter System or the Twist HT (High-Throughput) Universal Adapter System. The Twist Universal Adapter System consists of Twist Universal Adapters and Twist Unique Dual Indexed (UDI) Primers, whereas the Twist HT Universal Adapter System includes Twist Universal Adapters and HT UDI Primers. This library preparation protocol is optimized for use with Twist Target Enrichment Kits and should only be performed with reagents specified or their equivalents.
- User supplied materials and equipment
- General notes and precaution
- Guidelines for gDNA sample
- Twist Mechanical Fragmentation Library Preparation Workflow
DATA FILES
The number of commercially available Unique Dual Index (UDI) primer pairs remains limited, with few commercial offerings above 1,536 UDI pairs. In an era of ever-increasing sequencing capacity, this unnecessarily limits sample throughput and increases the cost per sample. To address these limitations, Twist Bioscience developed a new set of 3,072 UDIs that enables higher sample throughput in large experiments.
The design of large sets of barcodes can be difficult due to the exponential nature of barcode-barcode interactions. Twist’s design approach includes (i) sophisticated index design, (ii) subsetting index pairs into bins that are base and color channel balanced, and (iii) empirical validation of each index pair across multiple sequencing platforms. This approach enabled the generation of a large and performant set of UDI primers described here.
SKUs
- 101307 Twist Universal Adapter System TruSeq Compatible, 16 Samples
- 101308 Twist Universal Adapter System TruSeq Compatible, 96 Samples Plate A
- 101309 Twist Universal Adapter System TruSeq Compatible, 96 Samples Plate B
- 101310 Twist Universal Adapter System TruSeq Compatible, 96 Samples Plate C
- 101311 Twist Universal Adapter System TruSeq Compatible, 96 Samples Plate D
- 106390 Twist HT Universal Adapter System TruSeq Comp, 96W AAA01-04
- 106391 Twist HT Universal Adapter System TruSeq Comp, 96W AAA05-08
- 106392 Twist HT Universal Adapter System TruSeq Comp, 96W AAA09-12
- 106393 Twist HT Universal Adapter System TruSeq Comp, 96W AAA13-16
- 106396 Twist HT Universal Adapter System TruSeq Comp, 96W BBB01-04 (Coming soon!)
- 106398 Twist HT Universal Adapter System TruSeq Comp, 96W BBB05-08 (Coming soon!)
- 106399 Twist HT Universal Adapter System TruSeq Comp, 96W BBB09-12 (Coming soon!)
- 106400 Twist HT Universal Adapter System TruSeq Comp, 96W BBB13-16 (Coming soon!)
- 106401 Twist HT Universal Adapter System TruSeq Comp, 384W AAA01
- 106402 Twist HT Universal Adapter System TruSeq Comp, 384W AAA02
- 106403 Twist HT Universal Adapter System TruSeq Comp, 384W AAA03
- 106404 Twist HT Universal Adapter System TruSeq Comp, 384W AAA04
- 106405 Twist HT Universal Adapter System TruSeq Comp, 384W BBB01 (Coming soon!)
- 106406 Twist HT Universal Adapter System TruSeq Comp, 384W BBB02 (Coming soon!)
- 106407 Twist HT Universal Adapter System TruSeq Comp, 384W BBB03 (Coming soon!)
- 106408 Twist HT Universal Adapter System TruSeq Comp, 384W BBB04 (Coming soon!)
The Twist Universal Adapter System is for research use only, and subject to additional use restrictions as set forth in Twist’s Supply Terms and Conditions.
* For research use only. Not for diagnostic procedures.
