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Streamline ultra-high-throughput library prep

Scale your whole genome sequencing or targeted next generation sequencing studies without the bottleneck of manual normalization or prohibitive costs. The Twist FlexPrep™ UHT Library Preparation Kit is engineered for labs performing large-scale population genetics, agrigenomics, and related applications. One of the key applications of this kit is as an NGS genotyping solution, with advantages over microarray technology. Through Normalization by Ligation™ (NBL) technology, this kit eliminates the need for time-consuming individual sample quantification, allowing you to quickly move from raw DNA to pooled, sequencer-ready libraries.

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Integrated sample normalization

Eliminate manual quantification steps. Achieve consistent library yields across a flexible 30–300 ng input range using Normalization by Ligation™ technology.

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Reduce cost per sample

Early-stage sample pooling significantly reduces plastic waste and reagent usage without sacrificing data quality.

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Massive multiplexing scale

Manage large projects with ease. Uniquely index up to 1,152 samples per kit, ensuring precise sample identification and high-quality data analysis for even the largest study sizes.

Who we serve

Built for every team on the lab-compute divide

Whether you're building the next foundation model or advancing a clinical lead, Twist has a path for you.

Population Genomics

Population cohorts at population scale without the prep time

Population genomics demands processing hundreds or thousands of samples with consistent library quality, minimal batch effects, and cost-effective reagent use. FlexPrep UHT was engineered to meet exactly these demands, letting you scale your cohorts, not your consumable budget.

  • Process 1,152 samples in a single sequencing run from one kit
  • Consistent library normalization eliminates batch-to-batch variation
  • 12X sample pooling after ligation combined with 96-plex capture drastically reduces hands-on-time and maximizes sequencer utilization
  • Compatible with both whole genome sequencing and targeted sequencing
Learn more

Agrigenomics

From tissue punch to genotype call in a streamlined workflow

Agricultural samples are variable, challenging, and time-sensitive. We’ve paired FlexPrep with a DNA extraction method to bring you FlexPrep Pure Ag which handles fresh, frozen, dried, and stabilized plant and animal tissue without compromising data quality.

  • Single-step DNA purification—no ethanol, no extra washes
  • Validated on bovine ear punch tissue (TSU-collected, frozen samples)
  • High gDNA yield ~50 ng/µl with A260/A280 & A260/230 ratios >1.8
  • Target enrichment panels designed for animal and plant genomes
  • Suitable for fresh, frozen, dried, or otherwise stabilized sample types
Learn more
Product data

The Twist FlexPrep™ UHT Library Preparation Kit takes a novel Normalization by Ligation™ approach, which eliminates the need for
upfront and intermediate sample quantitation, streamlining your sequencing workflow.

Twelve inline barcodes are included on the FlexPrep normalization adapters, which allow for an ultra-high throughput and automation-friendly multiplexing strategy. With the FlexPrep UHT Library Preparation Kit, sequencing throughput can be maximized by running up to 1,152 samples in a single sequencing run from one kit. This increased efficiency can translate to cost and consumables savings. 

The self-normalizing adapters found in the Twist FlexPrep UHT Library Preparation Kit supports mass inputs of 30 ng to 300 ng with minimal impact on uniformity and on insert size for whole genome sequencing (WGS) or target enrichment (TE) workflows.

FlexPrep UHT generates high-complexity libraries that have uniform coverage after target enrichment. In combination with Twist Custom Panels, FlexPrep offers tunable coverage of SNPs, k-mers, structural variants, and other genomic areas of interest. Enriched material was downsampled to an average of 75x coverage. Key target enrichment metrics from Picard are reported.

Video
Diagram comparing a traditional enzymatic fragmentation NGS library preparation workflow with the Twist FlexPrep™ UHT Library Prep Kit. The illustration shows how Normalization by Ligation™ eliminates upfront and intermediate sample quantitation, reducing workflow steps, minimizing reactions from 96 to 8 after ligation, and streamlining next-generation sequencing (NGS) library preparation for 96 samples.
Twist FlexPrep UHT Library Prep Kit workflow showing inline barcodes and UDI indexing strategy for automated high-throughput NGS library preparation and multiplexing of up to 1,152 samples per sequencing run.
Graph showing normalized NGS read depth across variable DNA input amounts using the Twist FlexPrep UHT Library Prep Kit, demonstrating consistent library yield from 30 ng to 300 ng DNA inputs.
Target enrichment sequencing results from the Twist FlexPrep UHT Library Prep Kit showing uniform coverage across genomic regions using Twist Custom Panels and variable DNA input amounts.

Integrated normalization eliminates steps

The Twist FlexPrep™ UHT Library Preparation Kit takes a novel Normalization by Ligation™ approach, which eliminates the need for
upfront and intermediate sample quantitation, streamlining your sequencing workflow.

Diagram comparing a traditional enzymatic fragmentation NGS library preparation workflow with the Twist FlexPrep™ UHT Library Prep Kit. The illustration shows how Normalization by Ligation™ eliminates upfront and intermediate sample quantitation, reducing workflow steps, minimizing reactions from 96 to 8 after ligation, and streamlining next-generation sequencing (NGS) library preparation for 96 samples.

Comparison of typical enzymatic fragmentation (EF) to Twist FlexPrep UHT Library Preparation Kit using 96 samples. Typical library preparation requires labor-intensive quantitation and dilution prior to fragmentation. Reaction footprint also scales linearly with the number of samples. The Twist FlexPrep UHT Library Preparation Kit self-normalizes during ligation, and leverages inline barcoded adapters to enable a high-throughput workflow by compressing every 12 reactions into one pooled reaction.

Purpose-built workflow to save resources without compromising quality

Twelve inline barcodes are included on the FlexPrep normalization adapters, which allow for an ultra-high throughput and automation-friendly multiplexing strategy. With the FlexPrep UHT Library Preparation Kit, sequencing throughput can be maximized by running up to 1,152 samples in a single sequencing run from one kit. This increased efficiency can translate to cost and consumables savings. 

Twist FlexPrep UHT Library Prep Kit workflow showing inline barcodes and UDI indexing strategy for automated high-throughput NGS library preparation and multiplexing of up to 1,152 samples per sequencing run.

Fragmentation and ligation reactions are prepared in one well per sample. The adapters used during ligation contain inline barcodes that allow for the pooling of all 12 wells in a row of a 96-well plate. Individual pools are prepared with indices (UDIs) added by PCR for pool-level demultiplexing. Sequencing throughput can be maximized by running up to 1,152 samples in a single sequencing run from one FlexPrep kit. This increased efficiency can translate to cost and consumables savings.

Normalized yield with FlexPrep UHT

The self-normalizing adapters found in the Twist FlexPrep UHT Library Preparation Kit supports mass inputs of 30 ng to 300 ng with minimal impact on uniformity and on insert size for whole genome sequencing (WGS) or target enrichment (TE) workflows.

Graph showing normalized NGS read depth across variable DNA input amounts using the Twist FlexPrep UHT Library Prep Kit, demonstrating consistent library yield from 30 ng to 300 ng DNA inputs.

NGS read depth normalization with variable DNA mass input. 96 libraries were prepared with human genomic DNA (gDNA) (NA12878) using the Twist FlexPrep UHT Library Preparation Kit and FlexPrep Target Enrichment Protocol as a 96-plex with a custom 800 kb panel before sequencing on an Illumina NextSeq 550. Eight library pools of 12 samples each were generated using variable mass input ranging between 30 ng to 300 ng in each library pool. Percentage of total read counts identified to each library is calculated after unique dual index and inline barcode demultiplexing. Average with perfect normalization is estimated at 1.04% (100/96).

Tunable coverage for regions of interest

FlexPrep UHT generates high-complexity libraries that have uniform coverage after target enrichment. In combination with Twist Custom Panels, FlexPrep offers tunable coverage of SNPs, k-mers, structural variants, and other genomic areas of interest. Enriched material was downsampled to an average of 75x coverage. Key target enrichment metrics from Picard are reported.

Target enrichment sequencing results from the Twist FlexPrep UHT Library Prep Kit showing uniform coverage across genomic regions using Twist Custom Panels and variable DNA input amounts.

Metrics from a 96-plex enrichment using the Twist FlexPrep UHT Library Preparation Kit and FlexPrep Target Enrichment Protocol. A single 96-plex target enrichment was performed with an 800 kb panel with gDNA mass input ranging between 30 ng to 300 ng into library generation. The enriched material was sequenced on an Illumina NextSeq 550 and downsampled to an average of 75x coverage. Key target enrichment metrics from Picard are reported above.

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Diagram comparing a traditional enzymatic fragmentation NGS library preparation workflow with the Twist FlexPrep™ UHT Library Prep Kit. The illustration shows how Normalization by Ligation™ eliminates upfront and intermediate sample quantitation, reducing workflow steps, minimizing reactions from 96 to 8 after ligation, and streamlining next-generation sequencing (NGS) library preparation for 96 samples.

Integrated normalization eliminates steps

The Twist FlexPrep™ UHT Library Preparation Kit takes a novel Normalization by Ligation™ approach, which eliminates the need for
upfront and intermediate sample quantitation, streamlining your sequencing workflow.

Twist FlexPrep UHT Library Prep Kit workflow showing inline barcodes and UDI indexing strategy for automated high-throughput NGS library preparation and multiplexing of up to 1,152 samples per sequencing run.

Purpose-built workflow to save resources without compromising quality

Twelve inline barcodes are included on the FlexPrep normalization adapters, which allow for an ultra-high throughput and automation-friendly multiplexing strategy. With the FlexPrep UHT Library Preparation Kit, sequencing throughput can be maximized by running up to 1,152 samples in a single sequencing run from one kit. This increased efficiency can translate to cost and consumables savings. 

Graph showing normalized NGS read depth across variable DNA input amounts using the Twist FlexPrep UHT Library Prep Kit, demonstrating consistent library yield from 30 ng to 300 ng DNA inputs.

Normalized yield with FlexPrep UHT

The self-normalizing adapters found in the Twist FlexPrep UHT Library Preparation Kit supports mass inputs of 30 ng to 300 ng with minimal impact on uniformity and on insert size for whole genome sequencing (WGS) or target enrichment (TE) workflows.

Target enrichment sequencing results from the Twist FlexPrep UHT Library Prep Kit showing uniform coverage across genomic regions using Twist Custom Panels and variable DNA input amounts.

Tunable coverage for regions of interest

FlexPrep UHT generates high-complexity libraries that have uniform coverage after target enrichment. In combination with Twist Custom Panels, FlexPrep offers tunable coverage of SNPs, k-mers, structural variants, and other genomic areas of interest. Enriched material was downsampled to an average of 75x coverage. Key target enrichment metrics from Picard are reported.

Find your FlexPrep formulation

Four kit configurations cover every workflow from standard population genomics to agricultural field samples with integrated target enrichment.

Features

Standard Library Preparation

FlexPrep UHT Library Prep Kit

Library Preparation with Target Capture/Enrichment

FlexPrep UHT Library Prep & Enrichment Kit

FlexPrep UHT Library Prep with DNA extraction optimized for Ag

FlexPrep UHT Pure Ag DNA Library Prep Kit

Livestock-optimized

FlexPrep UHT Bovine LP & Enrichment Kit

Primacy audience

Population genomics, human genetics

Any species with custom TE panel

Agrigenomics, plant & animal research

Cattle & Livestock Breeding Programs

DNA purification module

User-supplied

User-supplied

✓

User-supplied

Normalization by Ligation™

✓

✓

✓

✓

Inline barcoding (12-plex)

✓

✓

✓

✓

Target enrichment/hybridization

-

✓

Optional + Hyb Kit

✓

Ag-specific workflow validation

-

-

-

✓

96-plex capture ready

-

✓

With Hyb Add-on

✓

Max throughput (1 kit)

192 or 1,152 samples

192 or 1,152 samples

192 or 1,152 samples

192 or 1,152 samples

Applications

Large-scale human or animal cohort studies where DNA is already extracted

Targeted resequencing of custom genomic regions across any species

Field-collected agricultural samples needing an end-to-end solution

Optimized target enrichment alternative to array-based genotyping for high-throughput bovine genomics

Optimized target enrichment alternative to array-based genotyping for high-throughput bovine genomicsOptimized target enrichment alternative to array-based genotyping for high-throughput bovine genomicsOptimized target enrichment alternative to array-based genotyping for high-throughput bovine genomicsOptimized target enrichment alternative to array-based genotyping for high-throughput bovine genomicsOptimized target enrichment alternative to array-based genotyping for high-throughput bovine genomicsOptimized target enrichment alternative to array-based genotyping for high-throughput bovine genomicsOptimized target enrichment alternative to array-based genotyping for high-throughput bovine genomicsOptimized target enrichment alternative to array-based genotyping for high-throughput bovine genomics.

Catalog numbers

109220 (192 samples)
109224 (1152 samples)

109223 (192 samples)
109226 (1152 samples)

127857 (192 samples)
127859 (1152 samples)

127858 (192 samples)
127861 (1152 samples)

Order now Contact Sales Rep
Order now Contact Sales Rep
Order now Contact Sales Rep
Order now Contact Sales Rep

Standard Library Preparation

FlexPrep UHT Library Prep Kit

Primacy audience

Population genomics, human genetics

DNA purification module

User-supplied

Normalization by Ligation™

✓

Inline barcoding (12-plex)

✓

Target enrichment/hybridization

-

Ag-specific workflow validation

-

96-plex capture ready

-

Max throughput (1 kit)

192 or 1,152 samples

Applications

Large-scale human or animal cohort studies where DNA is already extracted

Catalog numbers

109220 (192 samples)
109224 (1152 samples)
Order now Contact Sales Rep

Library Preparation with Target Capture/Enrichment

FlexPrep UHT Library Prep & Enrichment Kit

Primacy audience

Any species with custom TE panel

DNA purification module

User-supplied

Normalization by Ligation™

✓

Inline barcoding (12-plex)

✓

Target enrichment/hybridization

✓

Ag-specific workflow validation

-

96-plex capture ready

✓

Max throughput (1 kit)

192 or 1,152 samples

Applications

Targeted resequencing of custom genomic regions across any species

Catalog numbers

109223 (192 samples)
109226 (1152 samples)
Order now Contact Sales Rep

FlexPrep UHT Library Prep with DNA extraction optimized for Ag

FlexPrep UHT Pure Ag DNA Library Prep Kit

Primacy audience

Any species with custom TE panel

DNA purification module

User-supplied

Normalization by Ligation™

✓

Inline barcoding (12-plex)

✓

Target enrichment/hybridization

✓

Ag-specific workflow validation

-

96-plex capture ready

✓

Max throughput (1 kit)

192 or 1,152 samples

Applications

Targeted resequencing of custom genomic regions across any species

Catalog numbers

109223 (192 samples)
109226 (1152 samples)
Order now Contact Sales Rep

Livestock-optimized

FlexPrep UHT Bovine LP & Enrichment Kit

Primacy audience

Agrigenomics, plant & animal research

DNA purification module

✓

Normalization by Ligation™

✓

Inline barcoding (12-plex)

✓

Target enrichment/hybridization

Optional + Hyb Kit

Ag-specific workflow validation

-

96-plex capture ready

With Hyb Add-on

Max throughput (1 kit)

192 or 1,152 samples

Applications

Field-collected agricultural samples needing an end-to-end solution

Catalog numbers

127857 (192 samples)
127859 (1152 samples)
Order now Contact Sales Rep

Sequencing Partnerships

NGS library preparation and sequencing compatibility

The FlexPrep™ UHT Library Preparation Kit is designed for compatibility with leading next-generation sequencing platforms, including Illumina, Ultima, and Element sequencing systems. Build streamlined library preparation workflows across compatible platforms, or contact a Twist scientist to discuss customized sequencing workflows.


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FlexPrep sequencing compatibility partner logos

Frequently Asked Questions

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Normalization by Ligation™ is Twist’s proprietary technology to eliminate upfront normalization and intermediate quantitation, which allows work with sample batches using a wide range of inputs (30–300 ng).

Gencove data and internal bovine ear clip data are available. At launch, we support Human and Bovine. Contact Customer Support to access this data.

The recommended input amount is 30 ng - 300 ng, which should produce ≥ 1000 ng of final library with fragment sizes between 400-450 bp. Fragmentation size is dependent upon fragmentation time. For details, please refer to the Twist FlexPrep UHT Library Prep Protocol.

The Twist FlexPrep UHT Library Preparation Kit is compatible with DNA of different quality, where the kit has been tested with FFPE of various quality and degraded DNA. Contact Customer Support if you have additional questions

Yes, the kit can handle units of 12 samples, up to 192 total samples. The kits are configured to handle up to 192 and 1152 samples in each batch. If you would like to process less than 192 and 1152 samples, respectively, we recommend filling the remainder of the plate with sample replicates, which would offer even greater confidence in sequencing results.

Intended Use: For Research Use Only. Not for use in diagnostic procedures.

Performance & Normalization Disclaimer: Normalization by Ligation™ technology and library yields are validated within recommended gDNA mass inputs (30 ng to 300 ng). Individual results, sequencing coverage, and yield uniformity may vary based on sample quality, DNA integrity, operator execution, and platform selection.

Trademarks & Partner Disclosures: FlexPrep™ and Normalization by Ligation™ are trademarks of Twist Bioscience Corporation. Illumina® and NextSeq® are registered trademarks of Illumina, Inc. Element Biosciences®, Trinity™, and Trinity Freestyle™ are trademarks or registered trademarks of Element Biosciences, Inc. Gencove®, Curio Genomics™, Ultima Genomics®, and Picard are properties of their respective owners. Reference to third-party providers or platforms does not constitute or imply endorsement, sponsorship, or exclusive partnership unless expressly stated.

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