EGFR Plus Pathway Accel-Amplicon® Panel
All-in-One Amplicon SolutionUser Manual Brochure
The Accel-Amplicon® EGFR Pathway Panel offers contiguous coverage of EGFR and hotspot coverage of BRAF, KRAS, and NRAS, using a 17-amplicon design to generate multiplex libraries compatible with Illumina sequencing platforms. The Accel-Amplicon EGFR Pathway Panel is compatible with short DNA fragments from both FFPE and cfDNA samples.
This is especially well-suited for research focused on detecting clinically-relevant allele variants in DNA from circulating tumor cells (CTCs). This product is a complete kit that includes all elements necessary for generating ready-to-sequence libraries, including primer pairs and indexed sequencing adapters.
- Single-tube assay
- Ready-to-sequence libraries in 2 hours
- Inputs as low as 10 ng
- Average amplicon size of 136 bp for compatibility with cfDNA and FFPE
- Limit of detection as low as 1%
- On-target specificity and coverage uniformity > 95%
- Leverages the high fidelity performance of the Illumina platform
- Complete library generation in a single kit
The Accel-Amplicon workflow uses a fast, single-tube approach consisting of a 90-minute target enrichment amplification step and a 10-minute adapter ligation step, yielding a 2-hour start-to-finish procedure.
The Accel-Amplicon EGFR Pathway Panel offers high sensitivity variant detection from input amounts from 10-25 ng. The kit utilizes Illumina-compatible dual-indexed adapter sequences and has been validated on the MiSeq® and HiSeq® platforms.
*As quantified by qPCR. Qubit® represents amplifiable DNA content more accurately than NanoDrop®, however is still not as accurate as the qPCR assay. For sample types with more consistent high quality DNA including whole blood, fresh frozen samples, and cultured cells, quantification by Qubit is a reliable indicator of amplifiable content.
Genes Represented in the EGFR Pathway Panel and Number of Amplicons:
The Accel-Amplicon EGFR Pathway Panel includes hotspot loci and regions of contiguous coverage for EGFR. The table depicts the genes represented, followed by the number of amplicons for each gene. Contiguous, overlapping coverage is included for EGFR (highlighted in blue). Hotspot coverage is included for BRAF, KRAS, and NRAS.:
The data below demonstrate the performance of the Accel-Amplicon EGFR Pathway Panel reference DNA on Illumina Platform.
The Accel-Amplicon EGFR Pathway Panel consistently detected validated variants at the expected frequency in replicates from 10 ng of the Horizon Diagnostics Quantitative Multiplex DNA Reference Standards HD701. The variants were called by LoFreq 2.1.1 (Genome Institute of Singapore) and GATK HaplotypeCaller (Broad Institute). When examining uncommon variants between the 10 replicates, the majority of background variants were present at less than 0.6%. No sporadic variants greater than 0.6% were detected.
Sequencing data files (FASTQ, BED, BAM, and VCF) for this amplicon panel are available on request.