The HEAT-Seq Ultra Oncology HotSpot Panel has been discontinued. Our new KAPA Target Enrichment Portfolio is now available and contains our new KAPA HyperExome, as well as our new custom probes, KAPA HyperChoice and KAPA HyperExplore. For additional questions, please contact your regional sales representative.
The HEAT-Seq® Ultra Oncology HotSpot 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 635 probes, and has a capture target of approximately 30.5 Kb across previously described mutation hotspots in 53 commonly targeted oncogenes and is ideal for detecting and characterizing low-frequency mutations. The panel helps with the detection of variants at 1% minor allele frequency (MAF) and below.
Benefits of Heat-Seq Oncology HotSpot Panel
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.
Target the mutation hotspots in oncogenes with one panel
UABL1 |
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 |
HEAT-Seq Ultra Oncology HotSpot Panel performance
The hotspot panel was tested using 100 ng of FFPE DNA. 24 samples were multiplexed on a single MiSeq lane using v3 chemistry. Data from three replicate experiments is shown. High on-target coverage, uniformity, and coverage rates were observed for each replicate.
The HEAT-Seq Ultra Oncology HotSpot Panel was tested using an FFPE control sample with known mutations at frequencies between 0.9% and 25%. Observed allele frequencies strongly matched the known frequencies, showing the sensitivity and accuracy of this panel..
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.
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.
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 |
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:
Demo Data
Demo data is available for evaluation. Please contact Roche representative for your country.
Research Use Only. Not for use in diagnostic procedures.
HEAT-SEQ is a trademark of Roche.