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Reliable, scalable cfDNA workflows

Generate high quality libraries from challenging cfDNA inputs with a streamlined workflow that delivers high conversion and sensitivity for rare variant detection. Designed for low input and fragmented samples, the Twist cfDNA Library Preparation Kit helps you uncover critical insights with confidence and efficiency.

Reliable

Maximize Rare Variant Detection

High conversion rate reliably detects rare variants (≤0.1% VAF). Support duplex sequencing workflows with the Twist UMI Adapter System to maximize detection sensitivity.

samples

Streamlined workflow for precious samples

Achieve robust performance, even with low (<1ng) sample input. Get sequence-ready libraries in 2 hours.

detection

Amplification without Bias

Get consistent coverage across 5%–95% GC, preserving challenging genomic regions and difficult-to-capture targets. Consistent high yields from complex libraries — even at ≤100 pg DNA input. Call variants with confidence.

Product data

High fidelity amplification with Twist TrueAmp Polymerase generates cfDNA libraries that have substitution error rates comparable to those of PCR-free cfDNA libraries across all analyzed base changes.

Twist cfDNA Library Preparation Kit delivers higher cfDNA library yields. Following library generation, samples were eluted in 20 μL of water and 1 µL was quantified with QubitTM dsDNA Broad Range Kit.

Twist cfDNA Library Prep and Hyb Mix Kit enables sensitive, accurate detection down to 0.25% VAF. Single-plex capture with a 50 kb oncology panel targets 217 SNPs. Missed variants are sites with <1 variant family over total detectable sites.

Twist cfDNA Library Prep and Hyb Mix Kit delivers higher library complexity with or without UMI deduplication. Single plex capture with a 50 kb oncology panel targets variant sites in cfDNA standard material using optimized TE workflows.

Video
cfDNA sequencing error rate comparison showing PCR-free vs TrueAmp amplification libraries with equivalent base substitution accuracy for low-frequency variant detection
cfDNA library preparation kit generating higher DNA yield measured by Qubit, improving NGS library prep efficiency and sequencing performance
Sensitive detection of low-frequency variants at 0.25% VAF using cfDNA library prep and hybridization capture with oncology SNP panel
High-complexity cfDNA sequencing libraries with and without UMI deduplication, improving unique read depth and variant detection accuracy

High Fidelity Amplification with PCR

High fidelity amplification with Twist TrueAmp Polymerase generates cfDNA libraries that have substitution error rates comparable to those of PCR-free cfDNA libraries across all analyzed base changes.

cfDNA sequencing error rate comparison showing PCR-free vs TrueAmp amplification libraries with equivalent base substitution accuracy for low-frequency variant detection

Figure 1 Comparable Base Substitution Error Rates with and without TrueAmp Amplification. The Twist cfDNA Library Preparation Kit was used to generate libraries with Twist cfDNA Pan-Cancer Reference Standard v2 as the input material. Twist Full Length Unique Dual Index (UDI) Adapters were used during ligation for the PCR-free condition and Twist Universal Adapters were used during ligation for the PCR condition. The PCR condition was amplified using 6 cycles with Twist TrueAmp Polymerase mix and UDI primers. The libraries were pooled, sequenced and analyzed. Comparable error rates are seen between PCR-free and TrueAmp libraries.

Higher Conversion Rate = Higher Yield

Twist cfDNA Library Preparation Kit delivers higher cfDNA library yields. Following library generation, samples were eluted in 20 μL of water and 1 µL was quantified with QubitTM dsDNA Broad Range Kit.

cfDNA library preparation kit generating higher DNA yield measured by Qubit, improving NGS library prep efficiency and sequencing performance

Figure 2. Twist cfDNA Library Preparation Kit delivers higher cfDNA library yields. Following library generation, samples were eluted in 20 uL of water and 1 uL was quantified with Qubit™ dsDNA Broad Range Kit.

Higher Sensitivity & Higher Accuracy

Twist cfDNA Library Prep and Hyb Mix Kit enables sensitive, accurate detection down to 0.25% VAF. Single-plex capture with a 50 kb oncology panel targets 217 SNPs. Missed variants are sites with <1 variant family over total detectable sites.

Sensitive detection of low-frequency variants at 0.25% VAF using cfDNA library prep and hybridization capture with oncology SNP panel

Figure 3. Twist cfDNA Library Preparation and Hyb Mix Kit enables sensitive and accurate detection of variants at 0.25% VAF. Libraries were single-plex captured with a custom 50kb oncology panel targeting 217 SNP variant sites in the Pan-Cancer standard material. Variants Missed is calculated by summing sites with less than 1 variant family divided by total detectable variant sites.

Improved Duplexing = Higher Complexity

Twist cfDNA Library Prep and Hyb Mix Kit delivers higher library complexity with or without UMI deduplication. Single plex capture with a 50 kb oncology panel targets variant sites in cfDNA standard material using optimized TE workflows.

High-complexity cfDNA sequencing libraries with and without UMI deduplication, improving unique read depth and variant detection accuracy

Figure 4. Twist cfDNA Library Preparation and Hyb Mix Kit delivers higher complexity with and without UMI deduplication.

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cfDNA sequencing error rate comparison showing PCR-free vs TrueAmp amplification libraries with equivalent base substitution accuracy for low-frequency variant detection

High Fidelity Amplification with PCR

High fidelity amplification with Twist TrueAmp Polymerase generates cfDNA libraries that have substitution error rates comparable to those of PCR-free cfDNA libraries across all analyzed base changes.

cfDNA library preparation kit generating higher DNA yield measured by Qubit, improving NGS library prep efficiency and sequencing performance

Higher Conversion Rate = Higher Yield

Twist cfDNA Library Preparation Kit delivers higher cfDNA library yields. Following library generation, samples were eluted in 20 μL of water and 1 µL was quantified with QubitTM dsDNA Broad Range Kit.

Sensitive detection of low-frequency variants at 0.25% VAF using cfDNA library prep and hybridization capture with oncology SNP panel

Higher Sensitivity & Higher Accuracy

Twist cfDNA Library Prep and Hyb Mix Kit enables sensitive, accurate detection down to 0.25% VAF. Single-plex capture with a 50 kb oncology panel targets 217 SNPs. Missed variants are sites with <1 variant family over total detectable sites.

High-complexity cfDNA sequencing libraries with and without UMI deduplication, improving unique read depth and variant detection accuracy

Improved Duplexing = Higher Complexity

Twist cfDNA Library Prep and Hyb Mix Kit delivers higher library complexity with or without UMI deduplication. Single plex capture with a 50 kb oncology panel targets variant sites in cfDNA standard material using optimized TE workflows.

cfDNA Library Prep Kit with TrueAmp Polymerase Mix Workflow

The cfDNA workflow leverages engineered GC balance and high-fidelity amplification. This design delivers high amplification yield while minimizing PCR-introduced errors that can compromise low-frequency variant detection. The result is uniform, high-quality libraries in a fast, 2-hour workflow. Aptamer-based hot-start control provides room-temperature stability for automated setup, while the single-tube 2× mastermix streamlines both WGS and Target Enrichment workflows.

cfDNA NGS workflow with optimized GC balance and high-fidelity PCR amplification delivering uniform libraries, reduced sequencing errors, and improved low-frequency variant detection