The Twist cfDNA Pan-Cancer Reference Standard v2 is a meticulously engineered tool tailored for genomics professionals developing and validating NGS-based liquid biopsy assays. This reference standard provides a reliable and reproducible resource for detecting clinically relevant variants, enabling researchers to confidently evaluate assay performance at every stage. By integrating a wild-type (WT) cell-free DNA (cfDNA) background with synthetic oligonucleotides carrying mutant alleles, the material delivers exceptional precision and sensitivity.
Researchers can leverage this standard to define critical analytical parameters, including the Limit of Detection (LoD) and Limit of Blank (LoB), ensuring robust assay calibration and quality control. Ideal for both early-stage assay development and ongoing process monitoring, the Twist cfDNA Pan-Cancer Reference Standard v2 empowers genomics teams to accelerate liquid biopsy innovation while maintaining rigorous accuracy and reproducibility.
458 unique naturally occurring cancer variants
132 clinically relevant variants
Covers 84 different genes involved in cancer
Background DNA derived from human cfDNA samples
DNA size profile and post-sequencing profile mimic native cfDNA
Seven individual VAF percentages to choose from
Digital Droplet PCR verification of VAF percentages
Convenient test set of all VAF percentages available
Fragment Profile Comparison: Twist vs. Competitors
Twist cfDNA Pan-Cancer Reference Standard v2 closely mimics the fragment size distribution of native cfDNA, including distinct mononucleosomal and dinucleosomal peaks. When compared to competitor reference materials standardized for peak height, Twist’s standards provide a more biologically accurate representation of circulating cell-free DNA, supporting realistic assay development and reliable performance benchmarking.
Schematic of Synthetic Variant DNA Design
Twist’s synthetic variant DNA is engineered to tile across each genomic locus with extensive overlap, ensuring high coverage and diverse fragment representation. This design captures the variant in multiple positions relative to the DNA termini, providing realistic fragment diversity and enabling more accurate assay performance assessment.
Background Mutation Rate Comparison
Background mutation rate measures the occurrence of non-germline variants in duplex consensus reads from target-enriched wild-type samples. Analysis across Twist cfDNA Pan-Cancer Reference Standard v2, native cfDNA, competitors, and Twist v1 shows that v2 closely matches the low background error rate of native cfDNA. Its superior fidelity—lower than both v1 and competitor standards—is achieved through a high-precision production process, ensuring more reliable and accurate assay benchmarking.
INDEL Size Distribution
The Twist cfDNA Pan-Cancer Reference Standard v2 includes a wide range of insertion and deletion (INDEL) sizes, from deletions of 30 bp to insertions of 15 bp. Visualized as a histogram (bin width = 1), this distribution allows researchers to comprehensively evaluate INDEL detection performance across diverse fragment lengths. Single nucleotide variants (SNVs) are not included in this visualization, highlighting the standard’s focus on structural variant representation and assay sensitivity testing.
Variant Calling in Target Enrichment Workflows
Twist cfDNA Pan-Cancer Reference Standards deliver highly accurate variant detection across a range of variant allele frequencies (VAFs). Using a custom panel targeting 215 SNVs and sequencing to 80,000× raw coverage before UMI deduplication, the observed mean VAFs closely match the intended levels. This precision allows researchers to confidently benchmark and validate their NGS-based assays, ensuring reliable performance across the full spectrum of variant frequencies.
Detect low freqency variants from cfDNA with unique molecular IDs (UMIs).
A scalable target enrichment solution for monitoring minimal residual disease.
User-designed target enrichment panels for targeted NGS.
107576: Twist cfDNA Pan-Cancer Reference Standard v2 Set, 300ng kit
107577: Twist cfDNA Pan-Cancer Reference Standard v2 VAF 0% (WT), 3 ug
107578: Twist cfDNA Pan-Cancer Reference Standard v2 VAF 0.1%, 3 ug
107579: Twist cfDNA Pan-Cancer Reference Standard v2 VAF 0.25%, 3 ug
107580: Twist cfDNA Pan-Cancer Reference Standard v2 VAF 0.5%, 3 ug
107581: Twist cfDNA Pan-Cancer Reference Standard v2 VAF 1%, 3 ug
107582: Twist cfDNA Pan-Cancer Reference Standard v2 VAF 2%, 3 ug
107583: Twist cfDNA Pan-Cancer Reference Standard v2 VAF 5%, 3 ug
107584: Twist cfDNA Pan-Cancer Reference Standard v2 VAF 0%, 600ng
107585: Twist cfDNA Pan-Cancer Reference Standard v2 VAF 0.1%, 600ng
107586: Twist cfDNA Pan-Cancer Reference Standard v2 VAF 0.25%,600ng
107587: Twist cfDNA Pan-Cancer Reference Standard v2 VAF 0.5%, 600ng
107588: Twist cfDNA Pan-Cancer Reference Standard v2 VAF 1%, 600ng
107589: Twist cfDNA Pan-Cancer Reference Standard v2 VAF 2%, 600ng
107590: Twist cfDNA Pan-Cancer Reference Standard v2 VAF 5%, 600ng
Clinical Genomics Manager - ANZ
& Country Manager - NZ
Raise confidence in variant detection with superior target enrichment solutions
Precision, uniformity, and flexibility for results you can trust
Synthetic RNA and DNA standards for assay development
Identify more hits and streamline screening with Twist's precise Variant Libraries
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