Accel-Amplicon™ Comprehensive TP53 Panel

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Multiplex Amplicons for
Illumina Platforms.

All-in-One Amplicon Solution
The Accel-Amplicon™ Comprehensive TP53 Panel offers comprehensive coverage of all coding regions of the TP53 gene, using a 21-amplicon design to generate multiplex libraries compatible with Illumina sequencing platforms. Accel-Amplicon Comprehensive TP53 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.

Features:

  • Single-tube assay
  • Ready-to-sequence libraries in 2 hours
  • Inputs as low as 10 ng

Benefits:

  • Average amplicon size of 140 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

Workflow:
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.

amplicon-workflow-100416The single-tube workflow includes two brief incubations to generate the multiplex amplicon targets and add a unique combination of Illumina-compatible indexed adapters, creating up to 96 uniquely-indexed libraries for multiplexing on a single sequencing run.

Product Specifications:

Decrease Input, Not Sensitivity

Like the Accel-Amplicon 56G Oncology Panel v2, the Accel-Amplicon Comprehensive TP53 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.

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*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.
†Requires custom indexing option (please inquire).

Comprehensive Coverage of TP53

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Coverage of all coding regions of the TP53 gene splicing isoforms by the Accel-Amplicon Comprehensive TP53 Panel are represented in a Sashimi plot (IGV; Broad institute).

Performance Data:

Performance on the Illumina Platform

The data below demonstrate the performance of the Accel-Amplicon Comprehensive TP53 Panel on matched tumor-normal samples.

Variant Calling by Accel-Amplicon Comprehensive TP53 Panel
amplicon_table_variant-calling

The Accel-Amplicon Comprehensive TP53 Panel was compared with an Ion AmpliSeq Panel on cervical tumor FFPE samples and matched normal blood reference samples. 10 ng input DNA was used per sample and the libraries were sequenced to an average depth of 2000X on an Illumina MiSeq. The AmpliSeq assay called mutations across TP53 coding exons in 15 tumor samples. Somatic mutations were called using LoFreq 2.1.1 (Genome Institute of Singapore) and GATK HaplotypeCaller (Broad Institute). The discrepant calls were subsequently determined to be AmpliSeq false positives (indicated in red). Samples and AmpliSeq data were obtained from the National Cancer Institute (NCI).

Resources:

Protocols & Tools:

Product Overview:

Technical Notes:

Ordering:

Cat # AL-53048:  Accel-Amplicon Comprehensive TP53 Panel (48 rxns)