Mimix™ Multiplex I, cfDNA Reference Standard Set

The Multiplex I cfDNA Reference Standards are highly-characterized, biologically-relevant reference materials used to assess the performance of cfDNA assays that detect somatic mutations, including quantitative PCR, next-generation sequencing, and other novel assays

High throughput platforms such as Next generation sequencing (NGS) and quantitative PCR (qPCR) platforms give researchers much greater power when profiling cfDNA samples. However, the use of these presents challenges surrounding sample variability, platform bias, and potential failure to detect biomarkers. Horizon’s cfDNA products are all derived from human cell lines, and are fragmented to an average size of 160 bp to closely resemble cfDNA extracted from human plasma.

This format ensures that they may applied to a wide-range of cfDNA NGS and qPCR assays including high-throughput platforms such as NGS and qPCR give researchers greater power when profiling cfDNA samples. Horizon has developed the cfDNA Reference Standard range to support the development and continued validation of cfDNA assays.

The Multiplex I cfDNA Reference Standard Set covers multiple engineered single nucleotide variants (SNVs/SNPs) with 8 mutations at 5%, 1%, and 0.1% allelic frequencies. Batch-specific copy number values measured via ddPCR are provided with each Set.

cfDNA Application note

With this product you are able to:

Analyze the sensitivity and specificity of your assay
Gain certainty of the limit of detection and limit of quantification
Optimize and validate new cancer panels and routinely monitor the performance of your assay

Technical data

Format: cfDNA

Genes covered: EGFR, KRAS, NRAS, PIK3CA

Allelic frequencies: 5%, 1%, 0.1% and 0% (100% wild type)

Buffer: Tris-EDTA (10mM Tris-HCl, 1mM EDTA), pH 8.0

Product information

Fragment size: 160 bp

Unit size: 350 ng per vial

Concentration: 20 ng/µl

Verified mutations

100% Multiplex I Wild Type cfDNA Reference Standard (Part No.: HD776)
Chromosome	Gene	Variant	Expected Allelic Frequency
7p12	EGFR	L858R	0.00%
7p12	EGFR	ΔE746 - A750	0.00%
7p12	EGFR	T790M	0.00%
7p12	EGFR	V769 - D770insASV	0.00%
12p12.1	KRAS	G12D	0.00%
1p13.2	NRAS	Q61K	0.00%
1p13.2	NRAS	A59T	0.00%
3q26.3	PIK3CA	E545K	0.00%

5% Multiplex I cfDNA Reference Standard (Part No.: HD777)
Chromosome	Gene	Variant	Expected Allelic Frequency
7p12	EGFR	L858R	5.0%
7p12	EGFR	ΔE746 - A750	5.0%
7p12	EGFR	T790M	5.0%
7p12	EGFR	V769 - D770insASV	5.0%
12p12.1	KRAS	G12D	6.3%
1p13.2	NRAS	Q61K	6.3%
1p13.2	NRAS	A59T	6.3%
3q26.3	PIK3CA	E545K	6.3%

1% Multiplex I cfDNA Reference Standard (Part No.: HD778)
Chromosome	Gene	Variant	Expected Allelic Frequency
7p12	EGFR	L858R	1.0%
7p12	EGFR	ΔE746 - A750	1.0%
7p12	EGFR	T790M	1.0%
7p12	EGFR	V769 - D770insASV	1.0%
12p12.1	KRAS	G12D	1.3%
1p13.2	NRAS	Q61K	1.3%
1p13.2	NRAS	A59T	1.3%
3q26.3	PIK3CA	E545K	1.3%

0.1% Multiplex I cfDNA Reference Standard (Part No.: HD779)
Chromosome	Gene	Variant	Expected Allelic Frequency
7p12	EGFR	L858R	0.10%
7p12	EGFR	ΔE746 - A750	0.10%
7p12	EGFR	T790M	0.10%
7p12	EGFR	V769 - D770insASV	0.10%
12p12.1	KRAS	G12D	0.13%
1p13.2	NRAS	Q61K	0.13%
1p13.2	NRAS	A59T	0.13%
3q26.3	PIK3CA	E545K	0.13%

Presence confirmed in parental cell line: EGFR G719S (20% allelic frequency)

Exome sequencing data: Download additional information on variants present in the parental cell lines

General information

Storage: 4°C

Expiration: See all product shelf life information

Quality control

Fragmentation size: D1000 DNA ScreenTape assay

Allelic frequency: Droplet Digital™ PCR

Quantification: Qubit dsDNA BR Assay (post-fragmentation)

Intended use: For assay developers and molecular diagnostic labs for routine performance monitoring of molecular biology assays. Research use only. Not for diagnostic procedures.