The AmoyDx® BRCA1 and BRCA2 Gene Mutation Detection Kit (Reversible Terminator Sequencing) is intended for qualitative detection of BRCA1 and BRCA2 variants using DNA isolated from peripheral whole blood, fresh-frozen tissue or formalin-fixed paraffin-embedded (FFPE) tissue samples. The kit provides full and uniform coverage of both BRCA1 and BRCA2 genes, covering all coding exons, exon/intron junctions, some introns and UTR regions. The kit allows the detection of single nucleotide variants (SNVs) and insertions and deletions (Indels) from tissue-derived DNA, and the detection of SNVs, Indels, and large rearrangements (LRs) from blood-derived DNA.
The kit is intended to be used by trained professionals in a laboratory environment.
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||Cat. No|
|AmoyDx® BRCA1 and BRCA2 Gene Mutation Detection Kit||24T||-20℃||8.06.27203X024I|