HANDLE Classic NGS Panel

HANDLE Classic NGS Panel
The AmoyDx^® HANDLE Classic NGS Panel, based on HANDLE technology, is intended for qualitative detection of single nucleotide variants (SNVs), InDels, fusions, copy number amplifications (CNAs) and microsatellite instability (MSI) from DNA and RNA extracted from formalin-fixed paraffin-embedded (FFPE) tissue samples by converting sample nucleic acids to sequence-able libraries. The kit is intended to be used to aid clinician to identify 36 gene mutation status for cancer patients, including  several novel biomarkers (e.g. NTRK1~3, FGFR1~3 genes). 

Technological Principles
The test kit is based on Halo-shape ANnealing and Defer-Ligation Enrichment system (HANDLE system) technology which is an improved Molecular Inversion Probe (MIP) technology to capture the target gene region. The unique molecular identifier (UID) is introduced to both ends of each DNA fragment, and traces back to the original template for error correction. The library construction time of HANDLE system is 6 hours with 1 hour hands-on time. If use RNA sample, a reverse transcription step should be performed before hybridization with probes.
The probe contains an extension arm and a ligation arm which are complementary to the target gene region. First, the extension arm and ligation arm are anchored to the target gene region, and the DNA is extended from the extension arm to the ligation arm by the function of DNA polymerase. Next, the nicks are connected with the ligase to generate the circular products. The remaining linear probes, single-strand and double-strand nucleic acid are digested using exonuclease. Finally, the universal PCR amplification is performed to enrich the target libraries.

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