μCaler HotSpot Panel v1.0

Catalog Number:

#1101402
#1101401

μCaler 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 μCaler Hybrid Capture System. Based on the novel μCaler targeted enrichment system, the μCaler HotSpot Panel v1.0 delivers improved capture efficiency, consistent and excellent capture performance, and sig- nificantly reduces experimental time, enabling more efficient sequencing and cost savings.

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Product details
Kit Content
FAQ
Ordering Information
Resources

Gene list

AKT1	ALK	APC	AR	ARAF	ATM	BRAF	CDKN2A	CHEK2	CTNNB1
DDR2	EGFR	ERBB2	ERBB3	ESR1	FBXW7	FGFR1	FGFR2	FGFR3	FGFR4
FLT3	GNA11	GNAQ	GNAS	HRAS	IDH1	IDH2	KIT	KRAS	MAP2K1
MAP2K2	MET	MTOR	NRAS	NTRK1	NTRK3	PDGFRA	PIK3CA	PTCH1	PTEN
RAF1	RET	ROS1	SF3B1	SMAD4	SMO	STK11	TP53	TSC1

Performance

Capture Performance

Fig 1. Capture performance of μCaler 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-plex) input was used, following the user manual of μCaler 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).

Variant analysis

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).

For research use only. Not for use in diagnostic procedures.

Catalog	Color of Tube Cap	Item	Volume	Package/Storage
1101402		μCaler HotSpot Panel v1.0, 16 rxn	40 μL	-20℃
1101401		μCaler HotSpot Panel v1.0, 96 rxn	210 μL	-20℃

Can RNA samples be directly used for library preparation with this kit?

No. This kit is compatible only with gDNA or cDNA as initial samples. For RNA samples, reverse transcription to generate cDNA is required prior to library preparation.

What is the recommended input range? How to handle inputs exceeding this range?

This kit supports 50-2,000 ng of gDNA or cDNA. For input amount exceeds 2,000 ng, split the sample into multiple reactions to maintain amplification efficiency.

What is the insert size of this kit? How to select sequencing read length?

The main peak of PCR products by using this kit is ~270 bp. PE150 sequencing is recommended for high-quality coverage.

What is the recommended sequencing data volume for IGTR immune repertoire analysis?

A minimum of 0.3 Gb is recommended to detect clones at 0.01% abundance with an input of 200 ng. Increase data volume to enhance sensitivity for low-frequency clones.

Is this kit suitable for minimal residual disease (MRD) monitoring?

Yes. This kit includes IG Primer Mix and TR Primer Mix with gene-specific primers provided in separate tubes, allowing flexible combination in a single amplification reaction. With its high sensitivity, the kit meets the requirements for MRD monitoring technology development and clinical applications, making it ideal for low-frequency variant detection scenarios.