cfDNA Library Prep Kit with TrueAmp Polymerase Mix

Products

Applications

resources

COMPANY

PCR-Free WGS Library Preparation Kit

Overview Product Resources Application

Buy Now

Overview
Resources
Application

Buy Now

Engineered Performance for 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.

Maximize Rare Variant Detection

High conversion rate reliably detects rare variants (≤0.1% VAF). High coverage libraries compatible with duplex sequencing workflows supported by the Twist UMI Adapter System.

Streamline Workflow of Precious Samples

Robust performance, even with low (<1ng) sample input Sequence-ready libraries in 2 hours.

Streamline Workflow of Precious Samples

Robust performance, even with low (<1ng) sample input Sequence-ready libraries in 2 hours.

Product data

High Fidelity Amplification with PCR

High Fidelity Amplification with PCR Free–Like Error Rates cfDNA libraries prepared with Twist TrueAmp Polymerase show substitution error rates that are effectively equivalent to PCR-free cfDNA libraries across all measured base changes.

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.

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.

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.

Low Input Proven with native cfDNA

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

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.

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.

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.

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

Figure 5. Twist cfDNA Library Preparation Kit delivers higher cfDNA library yields with native cfDNA samples. Following library generation, samples were eluted in 20 uL of water and 1 uL was quantified with Qubit™ dsDNA Broad Range Kit.

Engineered to Preserve Library Yield at Low Input

Twist PCR-Free WGS Library Prep delivers robust library yields and consistent library size even as DNA input is reduced. Engineered ligase chemistry drives efficient, uniform ligation, helping maintain sequencing ready libraries across input levels.

Figure 1. PCR-free library yield comparison across inputs. Final library concentration (nM) was measured following PCR-free library preparation using the indicated DNA inputs. Twist PCR-Free WGS Library Preparation generated higher library yields than competitor workflows at equivalent inputs and maintained strong performance at lower inputs.

Reliable Insert Size Control Across Inputs

Twist PCR-Free WGS Library Prep maintains consistent median insert sizes from 300 ng down to 37.5 ng DNA input. Minimal read overlap across samples ensures efficient sequencing and dependable genome wide coverage.

Figure 2. The kit produces consistently large inserts across a wide range of DNA samples. (A)Median Insert Size is represented for 300 ng, 75 ng, and 37.5 ng of NA12878 DNA sample input. (B) Percent Overlap (which measures how much paired-end sequencing reads redundantly cover the same DNA bases) is represented for 300 ng, 75 ng, and 37.5 ng of NA12878 sample input.

Controllable Fragment Lengths for More Efficient Sequencing

Twist TrueAmp Library Preparation Kits enable precise control of insert size across a broad range, allowing users to tune library preparation to the specific requirements of their sequencing workflows.

Figure 3. Tunability of Twist PCR-Free WGS Library Prep Kit.

A: Five electropherograms of NGS libraries generated using differing fragmentation times. 50 ng of high-quality gDNA was fragmented for various times at 32°C.

Minimized GC Bias for More Uniform Coverage

Across DNA inputs, Twist PCR-Free WGS Library Prep maintains consistent coverage across GC content.

Figure 4. The Twist PCR-Free WGS Library Preparation Kit provides more consistent coverage distribution across varying GC content. The kit shows minimized GC bias even at low inputs, thereby reducing the need for additional sequencing to achieve uniform coverage and better detection of variants in the high- and low-GC content regions.

Minimized GC Bias for More Uniform Coverage

Across DNA inputs, Twist PCR-Free WGS Library Prep maintains consistent coverage across GC content.