SNVs, InDels, LRs in protein coding regions, intron/exon boundaries, some introns and UTR regions of the BRCA1 and BRCA2 genes
The AmoyDx® BRCA Pro Panel is a next-generation sequencing (NGS) based in vitro diagnostic assay intended for qualitative detection and classification of single nucleotide variants (SNVs), insertions and deletions (InDels), and large rearrangements (LRs) in protein coding regions, intron/exon boundaries, some introns and UTR regions of the BRCA1 and BRCA2 genes, using DNA isolated from peripheral whole blood or formalin-fixed paraffin embedded (FFPE) tumour tissue specimens. The detection of large rearrangement deletions and duplications is available for blood-derived DNA only. The assay is intended to aid in identifying breast cancer or ovarian cancer patients with pathogenic, or likely pathogenic BRCA variants.
This assay is not automated and is for laboratory professional use only.
Principles of the Procedure
The test kit is based on the Halo-shape ANnealing and Defer-Ligation Enrichment (HANDLE) system technology which is an improved Molecular Inversion Probe (MIP) technology to capture the target gene region (Figure 1). During the library construction process, each individual DNA molecule is tagged with a unique molecular index (UMI) at both ends, which allows high sensitivity in variant detection by eliminating any library amplification and sequencing bias. The test kit uses DNA extracted from tissue or blood samples, and it offers a time saving protocol that can be completed within 5 hours, and requires just about 1 hour of hands-on time.
Figure 1 Principle of library construction (HANDLE system)
The probe contains an extension arm and a ligation arm which are complementary to the target gene region. Firstly, the probe anneals onto the DNA template of the target region. Secondly, the DNA is extended from the extension arm to the ligation arm with the help of DNA polymerase, then the nicks are repaired to generate the circular products with the help of DNA ligase. Next, the remaining linear probes, single-strand and double-strand DNA are digested with the help of enzyme exonuclease, and only the target circular DNA will be kept for PCR amplification. Finally, the universal PCR amplification is performed to enrich the target DNA, and the magnetic bead-based purification is performed to obtain the final library.
After quality control (QC), the qualified libraries could be sequenced on Illumina sequencing platform. The sequencing data can be analyzed by AmoyDx NGS data analysis system (ANDAS) to detect the genomic variants in the target region
To order, please contact email@example.com.
|Product Name||Size (test/kit)||Storage|
|AmoyDx® BRCA Pro Panel||24T||-20℃|