HotSpot Detection | ^Caler HotSpot Panel v1.0IntroductionliCaler HotSpot Panel v1.0 targets the“hotspot" regions of tumor mutation, involving 49 genes related to carcinogenesis and tumor suppressor, with probes covering approximately 25 Kb of the genome.This Panel is designed for use with the pCaler Hybrid Capture System. Based on the novel pCaler targeted enrichment system, the liCaler HotSpot Panel v1.0 delivers improved capture efficiency, consistent and excellent capture performance, and significantly reduces experimental time, enabling more efficient sequencing and cost savings.Gene list AKT1 ALK GNAQ GNAS MTOR NRAS Mappability On-target 0.2x mean 0.5x mean depth depth Depth of coverage (mean = 100) Fig 1. Capture performance of ^Caler HotSpot Panel v1.0. LDT OncoOne Pan-tumor gDNA Standard (LDT Bioscience, LDT900) was used for pre-library preparation with NadPrep® EZ DNA Library Preparation Kit v2 paired with NadPrep® Universal Stubby Adapter (UDI) Module; 500 ng/pre-library (1-piex) input was used, following the user manual of liCaler hybrid capture. Sequencing was performed using Illumina Novaseq 6000, PE150. The BWA was used for alignment to the reference genome hg38 and On-target rate was calculated by the number of reads. A. Mappability, On-target rate and Target covered; B. Coverage uniformity. Note: OncoOne Pan-cancer gDNA Standard (LDT Bioscience, LDT900) include a negative control (LDT Bioscience, LDT898) and a positive control (LDT Bioscience, LDT899). Sequencing platform: Illumina Novaseq 6000,PE150.

Allele frequency Observed allele frequency (%) Observed allele frequency (%) Variant analysis Allele frequency for reference (%) Fig 2. Consistency of the allele frequency in μCaler HotSpot Panel v1.0 capture data with the reference frequency of the standards. Pre-libraries were prepared using the NadPrep EZ DNA Library Preparation Kit v2, μCaler HotSpot Panel v1.0 was used to complete hybridization capture. Sequencing was performed using Illumina Novaseq 6000, PE150. Note: The samples were: OncoOne Pan-cancer gDNA Standard (LDT Bioscience, LDT900). Ordering Information Product μCaler HotSpot...