• My account
  • Cart

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, PI3KCA

Cosmics

1,500+

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); download additional information on variants present in the parental cell lines:

Exome Sequencing Data

General Information

Storage

4°C

Expiry

See all product shelf life information

Cell Line Background

RKO/SW48

Quality Control

Fragmentation Size

D1000 DNA ScreenTape assay

Allelic Frequency

Droplet Digital™ PCR

Quantification

Qubit dsDNA BR Assay (post-fragmentation)