HEAT-Seq Ultra Oncology HotSpot Panel

Product Overview

The HEAT-Seq Ultra Oncology HotSpot Panel targets previously described mutation HotSpots in 53 oncogenes, and is ideal for detecting and characterizing mutations at low frequency.

ABL1

EZH2

JAK3

PTEN

AKT1

FBXW7

KDR

PTPN11

ALK

FGFR1

KIT

RB1

APC

FGFR2

KRAS

RET

ATM

FGFR3

MAP2K1

SMAD4

BRAF

FLT3

MET

SMARCB1

BRCA1

GNA11

MLH1

SMO

CDH1

GNAQ

MSH2

SRC

CDKN2A

GNAS

NF1

STK11

CSF1R

HNF1A

NOTCH1

TP53

CTNNB1

HRAS

NPM1

VHL

EGFR

IDH1

NRAS

 

ERBB2

IDH2

PDGFRA

 

ERBB4

JAK2

PIK3CA

 

Table 1. HEAT-Seq Ultra Oncology Panel genes

Benefits

  • Focus on previously-described mutational hot spots: The Oncology HotSpot panel allows you to further focus your sequencing resources with a smaller overall design size, simplifying analysis and enabling greater throughput.
  • Gain greater clarity into sample complexity: Detect variants at 5% MAF using this HotSpot panel. Have greater confidence in your discovery process and see through library preparation bias. Molecular barcoding made possible by the UIDs present in the HEAT-Seq probes allow you to detect and remove duplicates that are a result of PCR. This means you can confidently discern between a biological replicate of significance and a technological artifact.                                       
  • Maximize variant detection over a wide range of allelic frequencies: Leverage the UIDs to enable confident variant detection without the need for deep sequencing
  • Experience a simple workflow: Using HEAT-Seq accessory reagents, generate sequencing-ready DNA from sample DNA in less than a day with just two hours of hands-on time.
  • Remove false positives for greater confidence: Due to the stringent removal of amplification duplicates and process artifacts, sequencing and other errors are minimized. 

Product Performance

The HEAT-Seq Ultra Oncology HotSpot Panel provides uniform and complete coverage of the capture target.

HSQ HotSpot truq qmrs

Figure 1: HEAT-Seq Ultra Oncology HotSpot Panel performance
The HEAT-Seq Ultra Oncology HotSpot panel was tested using the Formalin Compromised Quantitative Multiplex Reference Standard and Tru-Q 4 Reference Standard (HD-C749 & HD731, Horizon Diagnostics). Each sample data set was subsampled to 500,000 reads. Average coverage depth from all sample was 360X.

HSQ HotSpot Accuracy

Figure 2: Accurate detection of variants using the HEAT-Seq Ultra Oncology HotSpot Panel
A plot of observed Vs. expected allele frequency using the HEAT-Seq Ultra Oncology HotSpot Panel and Formalin Compromised Quantitative Multiplex Reference Standard, HD-C749, Horizon Diagnostics.

HSQ HotSpot Figure Variant Depth

Figure 3: Coverage depth over regions of interest
The HEAT-Seq Ultra Oncology HotSpot panel was tested using the Formalin Compromised Quantitative Multiplex Reference Standard (HD-C749, Horizon Diagnostics). Using all available reads (3-5 M per sample), the panel provided coverage of between 250x and 1250x of known mutations.

 

 

HEAT-Seq Ultra Oncology HotSpot Panel

Illumina TruSeq Amplicon Cancer Panel

Thermo Fisher Ion AmpliSeq™ Cancer Panel v2

Average False-Positive Rate

0.014%

0.213%

3.071%

Range

0.0-0.2%

0.0%-0.4%

0.0-11.1%

Missed Calls

0

2

0

Figure 4: False-positive rate in detection of variants using various methods
The performance of the HEAT-Seq Ultra Oncology HotSpot Panel is compared to similar kits to assess allele frequency detection accuracy and data quality. The HEAT-Seq panel presents a similar or improved variant detection ability, but also presents an extremely low false-positive rate. The proprietary process of removing duplicates using molecular barcoding is the core of the HEAT-Seq system that enables not only accurate variant calls but also the confident removal of process induced errors that appear as false positives.

Design Files

The design and annotation files provide information about genomic regions covered by the capture probes and the genes included in these regions. These files were designed for use with the following Roche NimbleGen products:

  • HEAT-Seq Ultra Oncology HotSpot Panel, 24 Reactions (Catalog No. 07757069001)
  • HEAT-Seq Ultra Oncology HotSpot Panel, 96 Reactions (Catalog No. 07768613001)
 
 
 

Demo Data

Demo data is available for evaluation. Please contact your country Roche representative

Product Description

The HEAT-Seq Oncology Panel contains 635 probes, and has a capture target of approximately 30.5 Kb across 53 commonly targeted oncogenes.

Description Catalog Number Pack Size

HEAT-Seq Ultra Oncology HotSpot Panel  

07757069001

24 Reactions

HEAT-Seq Ultra Oncology HotSpot Panel  

07768613001

96 Reactions

Please Contact Us for ordering and questions

For life science research only. Not for use in diagnostic procedures. Unless otherwise stated in references, all data on file.

HEAT-Seq is a trademark of Roche. All other product names and trademarks are the property of their respective owners.

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