Order your ICEme kit

FOR RESEARCH USE ONLY

Improve Your Detection of Cancer Biomarkers

Improve mutation detection sensitivity 100-500 fold on any sequencing platform

Build your own ICEme Kit relevant to your cancer research
– choose from 17 Exons across BRAF, EGFR, KRAS, NRAS and PIK3CA.

Product Overview

ICEme Kit, uses Transgenomic’s exclusive Multiplexed ICE COLD-PCR (MX-ICP) technology, for the enrichment of mutations in the key cancer-related genes BRAF, EGFR, KRAS, NRAS & PIK3CA that are routinely analyzed in research on Colorectal, NSCLC, Melanoma & Breast cancer.

Performance

100 – ≥500 fold improvement of mutation detection sensitivity of all sequencing platforms (Sanger, NGS, ddPCR etc.)

Improves level of mutation detection (LOD) down to 0.01% or 5 copies in 50,000 genomic equivalents.

Enables detection of low level mutations for stratification of sub-populations for research projects.

Mutation Enrichment

Non allele-specific method that amplifies ALL mutations in the region under investigation, while suppressing wild-type alleles.

Multiplexed and customizable – you choose exons relevant to your cancer research.

Sample Flexibility

Compatible with DNA isolated from any sample type including liquid biopsies (plasma, urine, CSF) and tissues (fresh, frozen, FFPE, FNAs).

Ability to use liquid samples routinely will accelerate your research projects.

Platform Independent

Platform independent enabling analysis on any sequencing platforms (Sanger, NGS, ddPCR etc.).

ICEme Kit easily implementable into current workflows.

The first release of the ICEme Kit gives you the choice of selecting from 17 Exons:

  • BRAF Exon 11
  • BRAF Exon 15
  • EGFR Exon 12
  • EGFR Exon 18
  • EGFR Exon 19
  • EGFR Exon 20
  • EGFR Exon 21
  • KRAS Exon 2
  • KRAS Exon 3
  • KRAS Exon 4A
  • KRAS Exon 4B
  • NRAS Exon 2
  • NRAS Exon 3
  • NRAS Exon 4A
  • NRAS Exon 4B
  • PIK3CA Exon 9
  • PIK3CA Exon 20
Customize your own kit for your cancer research

For Research Use Only. Sale of this product is conditioned on the Limited Use License which you accept by purchasing this product.

The following are examples of Sanger sequencing electropherograms with and without mutation enrichment using MX-ICP. These mutations are reliably detected using the relevant MX-ICP Exon reactions. The Kit Positive Control was constructed by Horizon Discovery and consists of a mixture of digitally verified mutations. In the examples below, the “initial” percentage is the digitally verified percent mutation prior to MX-ICP and the “After MX-ICP” is the percent mutation determined after enrichment using the ICEme Kit.

cfDNAs from 5 plasma samples representing 4 different patients were analyzed for mutations using the ICEme Kit for BRAF Exon 15. Different mutations in this region can be detected following a single MX-ICP assay as well as the determination of Wild-Type sequences. The original mutation in the tumor is also listed. The tumor sample mutational analyses were performed by an independent third party.

150 ng of starting DNA containing 0.1 or 0.01% EGFR Exon 20 T790M Mutation Enriched using MX-ICP prior to ddPCR

150 ng of starting DNA containing 0.1 or 0.01% EGFR Exon 20 T790M Mutation Enriched using MX-ICP prior to ddPCR

ICEme Kit mutation enrichment was performed on cfDNA samples using NGS (MiSeq and Ion Torrent) Multiple mutations (across Exons 18 and 12) were enriched using MX-ICP compared with NGS alone.

The first release of the ICEme Kits gives you the choice of selecting from 17 Exons and Transgenomic will be continually adding more Exon targets to this panel. The ICEme Kit is a non-allele specific method that will amplify all mutations in the region under investigation. The possible number of alterations detectable (data from COSMIC) by the different kits is shown in the table below.

Tost, Jörg. The clinical potential of Enhanced-ice-COLD-PCR. Expert Review of Molecular Diagnostics. 2015 Dec 15; [Epub ahead of print]

Castellanos-Rizaldos, E., et al. COLD-PCR enriches low-level variant DNA sequences and increases the sensitivity of genetic testing. Methods Mol Biol. 2014 1102 623-39.

Castellanos-Rizaldos, E., et al. Single-tube, highly parallel mutation enrichment in cancer gene panels using temperature-tolerant-COLD-PCR. 2015 Jan;61(1):267–277.

Charbel, C., et al. NRAS mutation is the sole recurrent somatic mutation in large congenital melanocytic nevi. J Invest Dermatol. 2014 Apr 134(4) 1067-74.

How Kit, A., et al. Sensitive detection of KRAS mutations using enhanced-ice-COLD-PCR mutation enrichment and direct sequence identification. Hum Mutat. 2013 Nov 34(11) 1568-80.

How Kit, A., et al. Ultrasensitive detection and identification of BRAF V600 mutations in fresh frozen, FFPE, and plasma samples of melanoma patients by E-ice-COLD-PCR. Anal Bioanal Chem. 2014 Sep 406(22) 5513- 20.

How Kit, A. & Tost. J. Pyrosequencing® Based Identification of Low-Frequency Mutations Enriched Through Enhanced-ice-COLD-PCR. Methods Mol Biol. 2015 1315:83-101.

Milbury, CA ., et al. Ice-COLD-PCR enables rapid amplification and robust enrichment forlow-abundance unknown DNA mutations. Nucleic Acids Res. 2011 Jan; 39(1):e2.

Milbury,CA., et al. COLD-PCR enrichment of rare cancer mutations prior to targeted amplicon resequencing. Clin Chem. 2012 Mar; 58(3):580-9.

Sorich, M.J., et al. Extended RAS mutations and anti-EGFR monoclonal antibody survival benefit in metastatic colorectal cancer: a meta-analysis of randomized, controlled trials. Ann Oncol. 2015 Jan;26(1):13-21.

TBIO AACR 2016 Poster 1390

Very high sensitivity detection of K-RAS exon 2 mutations using Fast COLD-PCR. (AACR 2010)

Coupling conventional PCR and COLD-PCR amplified products, SURVEYOR® Nuclease and the Caliper LabChip® GX system for high sensitivity K-RAS mutation detection. (Caliper User Group 2010)

Development of a sensitive COLD-PCR method for the detection of EGFR mutations in DNA. (AACR 2011)

Use of BLOCker Sequencing (BLocking Oligonucleotide Cycle Sequencing) after Ice COLD-PCR for detection of K-RAS and BRAF mutations. (ASCO 2011)

Development of a sensitive COLD-PCR method for the detection of the EGFR mutations in plasma or serum. (ASCO 2011)

High sensitivity detection and robust enrichment for PIK3CA somatic mutations using ICE COLD-PCR technology. (ASCO 2011)

Very high sensitivity somatic mutation detection using ICE COLD-PCR and BLOCker sequencing. (ASHG 2011)

Very high sensitivity somatic mutation detection using ICE COLD-PCR and BLOCker sequencing. (ESHG 2011)

High sensitivity detection of PIK3CA somatic mutations using ICE COLD-PCR technology. (TRI-CON 2011)

Mutational analysis in circulating tumor cells (CTC): ScreenCell MB Filtration Unit and ICE COLD-PCR. (AACR 2012)

Ultrasensitive detection of mutations in the Androgen Receptor gene by ICE COLD-PCR. (AACR 2012)

Decipher genomic landscape of cancer mutations using ICE COLD-PCR and Ion Torrent Next-Generation sequencing technologies. (ADAPT 2013)

The use of improved and complete enrichment co-amplification at lower denaturation temperature (ICE COLD-PCR) method for the detection of EGFR and KRAS mutations from cell-free plasma DNA of non-small cell lung cancer (NSCLC) patients. (ASCO 2013)

Ultra-sensitive detection of BRAF V600E and G469A mutations by ICE COLD-PCR and BLOCKer sequencing. (Secon Diomarker Congress 2014)

BRAF and KRAS mutation testing in cell-free DNA and circulating tumor cells from blood of patients with metastatic cancers. (AACR 2014)

BRAF and KRAS mutation testing in plasma cell-free DNA with ICE COLD-PCR in patients with advanced cancers. (AACR 2014)

Mutant enrichment by ICE COLD-PCR prior to the Next-Generation sequencing enables high sensitivity and high throughput detection of cancer biomarkers in patient samples. (AACR 2014)

BRAF and KRAS mutation testing in plasma cell-free DNA with ICE COLD-PCR in patients with advanced cancers. (ASCO 2014)

Combination of DISSECTa and COLD-PCR technologies for ultra-sensitive detection of mutations using either Sanger or Next Generation sequencing methodologies. (ASCO 2014)

Mutant enrichment using ICE COLD-PCR paired to Next-Generation sequencing enables high sensitivity and high throughput detection of cancer biomarkers in patient samples. (Biomarker World Congress 2014)

Multiplexed ICE COLD-PCR: A mutation detection methodology for achieving sensitivities of <0.01% using either Sanger or NGS. (EORTC 2014)

Multiplexed ICE COLD-PCR (MX ICP): A highly sensitive mutation detection methodology that can achieve sensitivities <0.01% on Sanger and NGS on a single plate. (Next Generation Dx Conference 2014)

Utility of ICE COLD-PCR coupled to Sanger Sequencing or Next Generation Sequencing for increased mutation detection sensitivity during patient monitoring from circulating free DNA. (TRI-CON 2014)

Clinical Utility of expanded screening of the KRAS and NRAS genes for metastatic colorectal cancer – Evaluation of the CRC RAScan tumor test as a companion diagnostic to determine efficacy of anti-EGFR therapies. (ASCO 2015)

Specifications & Contents / Storage

Applications: Mutation enrichment, Biomarker discovery

Final Product Size: 95 – 159 bp

Form: Liquid

Reaction Type: Multiplexed PCR (MX PCR) and Multiplexed ICE COLD-PCR (MX-ICP)

Shipping Conditions: Dry Ice

Starting Material: DNA isolated from any sample type (liquid or tissue)

Storage: The kit should be stored between -18°C and -25°C in a constant temperature freezer until use.

Contents: Kits include MX PCR Primer Mix, MX-ICP Primers, RS-oligo, MX-ICP Positive Control, Phusion® Hot Start II DNA Polymerase, 5X GC Buffer for Phusion® HSII, JumpStart™ Taq DNA Polymerase, 10X JumpStart™ Reaction Buffer, dNTPs

Frequently Asked Questions

ICEme kit uses Transgenomic’s exclusive, ultra-sensitive, Multiplexed ICE COLD-PCR (MX-ICP) technology for the enrichment of mutations in the key cancer-related genes BRAF, EGFR, KRAS, NRAS and PIK3CA routinely analyzed in research on colorectal, NSCLC, melanoma and breast cancer. ICEme Kit enables use of tissue or liquid biopsy samples and provides up to ≥500-fold increase in mutation detection with LOD down to 0.01%
The first release of the ICEme Kit gives you the choice of selecting from 17 clinically actionable Exons / Mutations, across the key cancer-related genes BRAF, EGFR, KRAS, NRAS and PIK3CA, allowing you to build your own ICEme Kit customized to your cancer research. Transgenomic will be continually adding more Exon targets to this panel. The ultra-high sensitivity of the kits allows researchers to robustly detect low level mutations and stratify sub-populations for research projects
The ICEme Kit is platform independent allowing researchers to supercharge the sequencing platforms already present in their labs, including Sanger, NGS and ddPCR, to achieve greatly improved detection, up to a ≥500-fold increase, simply by adding the ICEme kits to the current workflow.

Customer Support

Contact Us By Phone or Submit Your Request by Online Form

NA & ROW: 1-800-362-3278 | EMEA: +44 141 892 8808

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Product
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Catalog Number

Unit Size
Reaction/Exon Cost
(USD)
List Price
(USD)
Place Order

ICEme 5 Exon Kit

716004-C

5 exons
24 reactions/exon

$30

$3,595

Select Exons

ICEme 10 Exon Kit

716005-C

10 exons
24 reactions/exon

$25

$5,895

Select Exons

ICEme 15 Exon Kit

716006-C

15 exons
24 reactions/exon

$24

$8,495

Select Exons