UNLEASHING THE POWER OF NEXT GENERATION SEQUENCING FOR ONCOLOGY
Product Information
Next generation sequencing (NGS) along with other multiplex platforms such as Sequenom give researchers much greater power when profiling samples. This presents new challenges, particularly with regard to variability between samples and potential failure to detect biomarkers. In response to this and with biomarker profiling of tumor samples in mind, Horizon Diagnostics (HDx) have developed a Quantitative Multiplex DNA Reference Standard. This is a highly annotated product, verified using digital PCR and containing a range of engineered cancer specific genotypes.
Key Features
- Precise allelic frequencies for major oncology targets
- Verified using digital PCR
- Recapitulates the complexity of tumor composition
- Renewable standard maintains assay consistency
- Quantitative standard drives confident interpretation of multiplex data
Pricing
| Pack Size | Pack Price | Price
per vial | Discount |
| 100ng |
$400.00 |
$400.00 |
-- |
| 200ng |
$750.00 |
$375.00 |
6.25% |
| 500ng |
$950.00 |
$190.00 |
52.5% |
| 1000ng |
$1500.00 |
$150.00 |
62.5% |
| 3000ng+ |
Contact Us |
|
|
| Pack Size | Pack Price | Price
per vial | Discount |
| 100ng |
€300.00 |
€300.00 |
-- |
| 200ng |
€562.50 |
€281.25 |
6.25% |
| 500ng |
€712.50 |
€142.50 |
52.5% |
| 1000ng |
€1125.00 |
€112.50 |
62.5% |
| 3000ng+ |
Contact Us |
|
|
| Pack Size | Pack Price | Price
per vial | Discount |
| 100ng |
£260.00 |
£260.00 |
-- |
| 200ng |
£487.50 |
£243.75 |
6.25% |
| 500ng |
£617.50 |
£123.50 |
52.5% |
| 1000ng |
£975.00 |
£97.50 |
62.5% |
| 3000ng+ |
Contact Us |
|
|
Background
Horizon Diagnostics derives value both from the GENESIS gene editing platform and the associated libraries of engineered isogenic cell lines. New isogenic cell lines continue to be developed to drive development of new quantitative reference standards. By tightly controlling the genetic identity and passage number of the various parental cell lines, Horizon Diagnostics have ensured that products are directly comparable, which fits well with the industry requirement for reducing variability.
Singleplex reference standards consist of an engineered heterozygous “mutant” cell line and a matched homozygous “wild type” cell line. Precise blending of isogenic cell lines allows creation of defined allelic frequencies as one would find in a tumor sample for example.
Multiplex reference standards take this concept further by combining as many as 20 different isogenic cell line pairs to give breadth of genetic coverage and depth in allelic frequency.
Allelic Frequencies
| Chromosome | Gene | Variant | Allelic Frequency |
| 7q34 |
BRAF |
V600E |
10.5% |
| 4q11-q12 |
cKIT |
D816V |
10.0% |
| 7p12 |
EGFR |
ΔE746 - A750 |
2.0% |
| 7p12 |
EGFR |
L858R |
3.0% |
| 7p12 |
EGFR |
T790M |
1.0% |
| 7p12 |
EGFR |
G719S |
24.5% |
| 12p12.1 |
KRAS |
G13D |
15.0% |
| 12p12.1 |
KRAS |
G12D |
6.0% |
| 12p12.1 |
NRAS |
Q61K |
12.5% |
| 12p12.1 |
PI3KCA |
H1047R |
17.5% |
| 12p12.1 |
PI3KCA |
E545K |
9.0%
|
Product Specification
| Property | Description |
| Purpose |
Analyzing and monitoring limit of detection of singleplex and multiplex assays
Research Use Only
|
| Number of Cell Lines Mixed |
8 |
| Cell Line Backgrounds |
HCT116, SW48, RKO |
| Vial Format |
100ng @ 5ng/μl or 200ng @ 50ng/μl |
| DNA Quality |
NO Transitions, Transversions, Insertions, Deletions, Ambiguous bases detected across the mutation of interest (~250 bp) |
| Stability |
Stable for >24 months from date of manufacture |
| Allelic Frequencies |
Verified using Digital PCR |
| MSDS |
 |
| Application Note: |
|
Please contact us if you have any questions: technical@horizondx.com
Horizon Diagnostics has worked with key partners to perform beta testing on the new product. The observed mutant frequency was verified by HDx using digital PCR and validated by Partner Organisations using the Ion Torrent™ Sequencer.
| Source | Horizon Dx | Partner A | Partner B | Partner C |
| Platform |
QX100 Droplet Digital PCR |
Custom Cancer Panel |
Ampliseq Cancer Hotspot v2 |
Ampliseq Cancer Hotspot v2 |
| Sequencing Depth |
N/A |
3000-4000x |
5000x |
2000x |
| Gene |
Mutation |
Observed Mutation |
| BRAF |
V600E |
10.2% |
9.9% |
9.1% |
10.3% |
| cKIT |
D816V |
10.4% |
10.0% |
11.0% |
10.1% |
| EGFR |
ΔE746-A750 |
2.0% |
2.3% |
Not Detected |
Not Detected |
| EGFR |
L858R |
2.7% |
2.7% |
2.1% |
2.4% |
| EGFR |
T790M |
0.9% |
0.8% |
Not Detected |
Not Detected |
| EGFR |
G719S |
24.4% |
23.7% |
23.1% |
24.8% |
| KRAS |
G13D |
16.1% |
16.3% |
12.35% |
15.5% |
| KRAS |
G12D |
5.0% |
5.2% |
Not Detected |
5.1% |
| NRAS |
Q61K |
12.8% |
9.0% |
12.7% |
12.6% |
| PIK3CA |
H1047R |
18.6% |
16.7% |
16.8% |
17.9% |
| PIK3CA |
E545K |
8.9% |
3.2% |
8.4% |
8.8% |
A review of the data supports the following observations:
- Ion Torrent™ data supports the product specification in most instances, agreeing broadly with the digital PCR data
- The PIK3CA E545K genotype is sometimes called incorrectly when assessed using Ion Torrent
- The inconsistent detection of the low frequency KRAS G12D, EGFR ΔE746-A750 and EGFR T790M mutations, even when using a common sequencing platform illustrates the need for a quantitative reference standard
- Sequencing depth, based on the limited data set, does not appear to limit or improve detection
Please contact us if you have any questions: technical@horizondx.com
Quality Control Summary
The following tests are run as standard to ensure the quality of our quantitative molecular reference standards:
| Test | Method |
| Allelic Frequency |
Droplet Digital PCR™ |
| Cell line ID |
Microsatellite profiling of source cell line |
| Genotype |
Sanger sequencing of locus specific PCR |
| Quality |
Agarose gel electrophoresis, Quantitatitive PCR of GAPDH locus |
| Quantification |
Quantifluor™ assay |
Please contact us if you have any questions: technical@horizondx.com
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