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Accel-Amplicon™ BRCA1 and BRCA2 Panel

AL52048

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.

Main features

  • Single-tube assay
  • Ready-to-sequence libraries in 2 hours
  • Inputs as low as 10 ng
  • Limit of detection as low as 1-5% for somatic mutations

More details

Description
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Description

Decrease input, not sensitivity

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).

Features

  • Single-tube assay
  • Ready-to-sequence libraries in 2 hours
  • Inputs as low as 10 ng
  • Average amplicon size of 148 bp for compatibility with FFPE and cfDNA
  • Limit of detection as low as 1-5% for somatic mutations
  • On-target specificity and coverage uniformity > 95%
  • Leverages the high fidelity performance of the Illumina platform
  • Complete library generation in a single kit

Comprehensive coverage of coding sequences of BRCA1

brca1_sashimi.png

Coverage of all coding exons of the BRCA1 gene by Accel-Amplicon BRCA1/2 Panel are represented in a Sashimi plot (IGV; Broad Institute).

Comprehensive coverage of coding sequences of BRCA2

Coverage of all coding exons of the BRCA2 gene by Accel-Amplicon BRCA1/2 Panel are represented in a Sashimi plot (IGV; Broad Institute).

Performance on the Illumina platform

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).

Specifications

FeatureSpecification
InputInput DNA required10-25 ng*
Sample typesFFPE, cfDNA, fresh frozen, gDNA
WorkflowTime required2 hours
Multiplexing on MiSeq v2 Nano at 5'000X average depth (germline)16
Multiplexing on MiSeq v2 at 5'000X average depth (somatic)24
DesignNumber of amplicons246
Amplicon sizeAverage 148 bp
Number of genes covered2
Total target size22.96 kb
PerformanceOn target percentage> 95%
Coverage uniformity at > 20% of mean> 95%
*As quantified by qPCR. Qubit® represents amplifiable DNA content more accurately than NanoDrop®, however it 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.

Resources