The HEAT-Seq® Oncology Panel is based on amplification-based enrichment employing an advanced version of Molecular Inversion Probe (MIP) technology and combines a fast and easy workflow with powerful SNV detection. The panel contains 5,144 probes, and has a capture target of approximately 245 Kb across 60 commonly targeted oncogenes and is ideal for new variant discovery and validation.

Benefits of Heat-Seq Oncology Panel

  • Simple workflow with sequencing-ready DNA in a single tube with just two hours of handson time
  • Detection and removal of PCR duplicates using molecular barcoding and the ability to confidently discern a biological replicate and a process artifact
  • Maximized variant detection over a wide range of allelic frequencies using molecular barcoding with UIDs without the need for deep sequencing
  • Minimized sequencing errors due to the stringent removal of false positives
  • Utilize HSQutils software, which is a separately sourced sequencing read mapper and variant caller for use in the analysis workflow

Heat-Seq Workflow

The final product of the workflow is a sequencing-ready amplicon that contains the target DNA sequence, a molecular barcode, and sample indices.

The entire workflow is completed in 8 hours with < 2 hours of hands-on time using a single tube. The protocol is easy to adopt and is also automation-friendly.

 

Product Highlights

Target the coding region of 60 oncogenes with one panel 

HEAT-Seq Oncology Panel genes

ABL1

EZH2

KDR

PDGFRA

AKT1

FBXW7

KIT

PIK3CA

ALK

FGFR1

KRAS

PTEN

APC

FGFR2

MAP2K1

PTPN11

ATM

FGFR3

MAP2k4

RB1

BRAF

FLT3

MET

RET

BRCA1

GNA11

MLH1

RUNX1

BRCA2

GNAQ

MPL

SMAD4

CDH1

GNAS

MSH2

SMARCB1

CDKN2A

HNF1A

MSH6

SMO

CSF1R

HRAS

NF1

SRC

CTNNB1

IDH1

NF2

STK11

EGFR

IDH2

NOTCH1

TERT

ERBB2

JAK2

NPM1

TP53

ERBB4

JAK3

NRAS

VHL

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Accurate, reliable and uniform performance

HEAT-Seq Oncology Panel performance

The HEAT -Seq Oncology Panel was tested using Coriell HapMap 12878 DNA. Accuracy, reliability and uniformity are demonstrated across many replicates. Each replicate was subsampled to 2 million reads, and yielded a mean coverage across the target of 163X. Uniformity is defined is the percentage of probes giving coverage that is greater than or equal to 20% of the mean.

Strong detection of known SNPs

HEAT-Seq Oncology Panel Variant Detection.

The HEAT-Seq Oncology Panel was used to measure allele frequency in a sample of formalin-compromised DNA (Quantitative Multiplex Reference Standard, HD-C749, Horizon Discovery) to simulate the effects of FFPE storage and extraction. Detection of variants at known frequencies between 0.9% and 25% is demonstrated with high correlation. Each replicate was tested using 250ng of DNA and data was subsampled to 2 million reads per sample.

Accurate detection of variants due to removal of duplicates and artifacts by UIDs

View data on HEAT-Seq oncology panel specificity and accurate detection of variants at a wide frequency range with zero total false positive rates.*

Chromosome Coordinate Expected Variant MAF Average Variant MAF Detected Total False Positives

chr1

115256529

12.5

6%

0

chr3

29443695

8.75

9%

0

chr3

178936091

17.5

15%

0

chr4

178952085

10

7%

0

chr7

55241707

24.5

24%

0

chr7

55259515

0.9

1%

0

chr7

140453136

2.8

2%

0

chr7

140453136

10.7

12%

0

chr12

80409488

15

12%

0

chr12

123279677

6.25

6%

0

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 products:

  • HEAT-Seq Oncology Panel, 24 Reactions (Catalog No. 07757042001)
  • HEAT-Seq Oncology Panel, 96 Reactions (Catalog No. 07768605001)

Download Design Files

Demo Data

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

*Data on file