Simplified Workflow for Ultra-High Throughput Sequencing
Twist’s FlexPrepTM UHT Library Preparation Kit offers an NGS alternative to traditional microarray-based technologies for population studies, agrigenomics and other ultra-high throughput applications. Combining Twist’s Normalization by LigationTM (NBL) technology with enzymatic fragmentation methods, the kit offers built-in sample normalization across a wide range of DNA inputs, reducing both cost and complexity of typical sample processing steps. Early sample barcoding enables downstream pooling of twelve separate samples into one reaction for a more streamlined and efficient workflow.
For research use only. Not for use in any diagnostic or clinical procedures.
*Based on a 8 well vs 96 well workflow
Discover Twist's approach to ultra-high throughput sequencing
FAQ
Simplified Workflow for Ultra-High Throughput Sequencing
Twist’s FlexPrepTM UHT Library Preparation Kit offers an NGS alternative to traditional microarray-based technologies for population studies, agrigenomics and other ultra-high throughput applications. Combining Twist’s Normalization by LigationTM (NBL) technology with enzymatic fragmentation methods, the kit offers built-in sample normalization across a wide range of DNA inputs, reducing both cost and complexity of typical sample processing steps. Early sample barcoding enables downstream pooling of twelve separate samples into one reaction for a more streamlined and efficient workflow.
This document provides more details and supporting data on performance.
Discover Twist's approach to ultra-high throughput sequencing
For research use only. Not for use in any diagnostic or clinical procedures.
*Based on a 8 well vs 96 well workflow
FAQ
The Twist FlexPrepTM UHT Library Preparation Kit takes a novel Normalization by LigationTM approach which eliminates the need for upfront and intermediate sample quantitation, streamlining your sequencing workflow.
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.
NGS read depth normalization with variable DNA mass input. 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).
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 using a 96-plex30 ng to 300 ng gDNA Mass Input into Library Preparation |
|
Metrics at 75x Raw Target Coverage |
Average +/- Standard Deviation |
Selected BasesMean Target CoverageChimerasFold-80 Base PenaltyCovered Bases at 10XCovered Bases at 20XCovered Bases at 0X |
79.22% +/- 0.54%32.74 +/- 4.781.31% +/- 0.38%1.416 +/- 0.04496.06% +/- 0.94%85.41% +/- 6.36%0.43% +/- 0.04% |
*Method used: 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. **Standard kits based on workflows which use enzymatic fragmentation followed by ligation and PCR ***All charts, figures, and graphs per Twist internal data September 2024. For research use only. Not for use in any diagnostic or clinical procedures.
The Twist FlexPrepTM UHT Library Preparation Kit takes a novel Normalization by LigationTM approach which eliminates the need for upfront and intermediate sample quantitation, streamlining your sequencing workflow.
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.
NGS read depth normalization with variable DNA mass input. 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).
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 using a 96-plex30 ng to 300 ng gDNA Mass Input into Library Preparation |
|
Metrics at 75x Raw Target Coverage |
Average +/- Standard Deviation |
Selected BasesMean Target CoverageChimerasFold-80 Base PenaltyCovered Bases at 10XCovered Bases at 20XCovered Bases at 0X |
79.22% +/- 0.54%32.74 +/- 4.781.31% +/- 0.38%1.416 +/- 0.04496.06% +/- 0.94%85.41% +/- 6.36%0.43% +/- 0.04% |
*Method used: 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. **Standard kits based on workflows which use enzymatic fragmentation followed by ligation and PCR ***All charts, figures, and graphs per Twist internal data September 2024. For research use only. Not for use in any diagnostic or clinical procedures.
109220
Twist FlexPrep™ UHT Library Preparation Kit, 192 Samples109223
Twist FlexPrep™ UHT LP and Hybridization Kit, 192 Samples109224
Twist FlexPrep™ UHT Library Preparation Kit, 1152 Samples109226
Twist FlexPrep™ UHT LP and Hybridization Kit, 1152 Samples109220
Twist FlexPrep™ UHT Library Preparation Kit, 192 Samples109223
Twist FlexPrep™ UHT LP and Hybridization Kit, 192 Samples109224
Twist FlexPrep™ UHT Library Preparation Kit, 1152 Samples109226
Twist FlexPrep™ UHT LP and Hybridization Kit, 1152 Samples
Product Sheet
Protocol
Guide/Guideline
Protocol
Twist FlexPrep UHT Library Preparation Kit with Enzymatic Fragmentation and Twist UDI Primers
