Quantitative Multiplex Reference Standard (gDNA)

The Quantitative Multiplex DNA Reference Standard is a highly-characterized, biologically-relevant quality control material used to assess the performance of NGS assays that detect somatic mutations.

Next generation sequencing (NGS) platforms give researchers much greater power when profiling tumor samples. However, the use of NGS presents challenges surrounding sample variability, platform bias, and potential failure to detect biomarkers. Horizon’s standards are appropriate for any NGS library preparation including whole-genome, whole-exome, custom capture and targeted amplicon panels. Horizon has developed this Quality-Seq (Q-Seq) NGS Reference Standard range to support the development and continued validation of Next Generation Sequencing platforms. The Quantitative Multiplex Reference Standard (QMRS) portfolio covers multiple endogenous SNPs, insertions and deletions. The QMRS includes 11 mutations at 0.8-24.5% allelic frequency in genomic DNA, FFPE and Formalin-Compromised DNA format. The multiple formats of the QMRS allow you to validate both the pre-analytical and analytical portion of your workflow using an isogenic reference standard. The FFPE format QMRS allows you to evaluate and optimize DNA extraction protocols without wasting precious patient samples. With the Formalin-Compromised format, the robustness of your NGS workflow may be tested to ensure that your library protocol and informatics pipeline can accommodate these highly degraded samples. The QMRS standard is our most popular multiplex standard as it provides both a breadth of formats, but also a large range of allelic frequencies and onco-relevant mutants.

With this product you are able to:

Analyze the specificity of your assay
Evaluate the robustness of your workflow with multiple formats (DNA , FFPE and Formalin-Compromised DNA)
Optimize and validate new cancer panels and routinely monitor the performance of your assay

Technical Data

Format: Genomic DNA

Genes Covered: BRAF, KIT, EGFR, KRAS, NRAS, PIK3CA

Allelic Frequencies: 1% - 24.5% for verified variants

Buffer: Tris-EDTA (10mM Tris-HCl, 1mM EDTA), pH 8.1

Product Information

Verified Mutations:

Chromosome	Gene	Variant	Expected Allelic Frequency, %
7q34	BRAF	V600E	10.50%
4q11-q12	cKIT	D816V	10.00%
7p12	EGFR	ΔE746 - A750	2.00%
7p12	EGFR	L858R	3.00%
7p12	EGFR	T790M	1.00%
7p12	EGFR	G719S	24.50%
12p12.1	KRAS	G13D	15.00%
12p12.1	KRAS	G12D	6.00%
1p13.2	NRAS	Q61K	12.50%
3q26.3	PIK3CA	H1047R	17.50%
3q26.3	PIK3CA	E545K	9.00%

Presence confirmed in parental cell line

Chromosome	Gene	Variant	Expected Allelic Frequency, %
2p23	ALK	P1543S	33%
1q25.2	ABL2	P986fs	8%
5q21-q22	APC	R2714C	33%
1p35.3	ARID1A	p.M1564fs*1	33.50%
13q12.3	BRCA2	K1691fs	33%
13q12.3	CDX2	V306fs	41.50%
22q13.2	EP300	K292fs	8%
4q31.3	FBXW7	S668fs	33.50%
8p12	FGFR1	P150L	8.50%
13q12	FLT3	V197A	11.50%
2q33.3	IDH1	S261L	10%
7q31	MET	L238fs	6.50%
3p21.3	MLH1	L323M	8.50%
17q11.2	NF1	Y628fs	7.50%
22q12.2	NF2	P275fs	8%
9q34.3	NOTCH1	P668S	31.50%
1q21-q22	NTRK1	5'UTR (1:156815754, GRCh38)	8.50%
4q12	PDGFRA	G426D	33.50%

Unit Size: 1 µg

Concentration: 50 ng/µl

General Information

Storage: 4˚C

Expiration: See all product shelf life information

Quality Control

Allelic Frequency: Droplet Digital PCR™

Genotype: Sanger sequencing of locus specific PCR

Quality: Agarose gel electrophoresis

Quantification: Spectrophotometry (A260)