Targeted sequencing for breast cancer genes - all-in-one amplicon solution
The Accel-Amplicon BRCA1 and BRCA2 Panel offers comprehensive coverage of the entire coding sequence of the BRCA1 and BRCA2 genes. Utilizing 246 amplicons with an average size of 148 bp, this panel generates targeted libraries compatible with Illumina® and Ion Torrent™ sequencing platforms. The unique design of the Accel-Amplicon BRCA1 and BRCA2 Panel enables compatibility with FFPE and cfDNA samples. This product is a complete kit that includes all components necessary for generating ready-to-sequence libraries, including primer pairs and indexed sequencing adapters.
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Like the Accel-Amplicon BRCA1, BRCA2, and PALB2 Panel, the Accel-Amplicon BRCA1 and BRCA2 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 MiniSeq® platforms. Barcoded adaptors for sequencing on Ion Torrent platforms are also available (please inquire).
Coverage of all coding exons of the BRCA1 gene by Accel-Amplicon BRCA1/2 Panel are represented in a Sashimi plot (IGV; Broad Institute).
Coverage of all coding exons of the BRCA2 gene by Accel-Amplicon BRCA1/2 Panel are represented in a Sashimi plot (IGV; Broad Institute).
The data below demonstrate the performance of the Accel-Amplicon BRCA1 and BRCA2 Panel on a variety of sample types.
The Accel-Amplicon™ BRCA1/2 Panel was used to prepare libraries from 10 ng inputs of high quality Coriell (NA12878, NA24143, NA24695, and NA19240) gDNA, Horizon Diagnostics Quantitative Multiplex DNA Reference Standards HD701 and FFPE samples subjected to 6, 24, or 48 hours of fixation. Sequencing was performed using MiniSeq® Reagents.
Variant calling by Accel-Amplicon Comprehensive BRCA1 and BRCA2 Panel
The Accel-Amplicon BRCA1/2 Panel consistently detected validated variants at the expected allele frequency (AF) in replicates from 10 ng of the Horizon Diagnostics Quantitative Multiplex DNA Reference Standards HD701 and Coriell (NA14684-21, NA14805-26 and NA14788-27). The variants were called by GATK HaplotypeCaller (Broad Institute).
|Input||Input DNA required||10-25 ng*|
|Sample types||FFPE, cfDNA, fresh frozen, gDNA|
|Workflow||Time required||2 hours|
|Multiplexing on MiSeq v2 Nano at 5'000X average depth (germline)||16|
|Multiplexing on MiSeq v2 at 5'000X average depth (somatic)||24|
|Design||Number of amplicons||246|
|Amplicon size||Average 148 bp|
|Number of genes covered||2|
|Total target size||22.96 kb|
|Performance||On target percentage||> 95%|
|Coverage uniformity at > 20% of mean||> 95%|