Product details

μCaler HyperMix Hybrid Capture Kit

Catalog Number:

#1105302 #1105301
#1105402 #1105401

μCaler HyperMix Hybrid Capture Kit is a super-premixed reagent optimized for targeted enrichment of μCaler Panels/Probes (20-100 nt non-isolength) and hybrid capture of various DNA libraries. Featuring the HyperMix design, the hybrid reaction can be prepared simply by adding the probe and library template, streamlining workflow while improving experimental efficiency and operational consistency.

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Feature

Super-premixed Design for a Simpler and More Efficient Workflow

● Ready-to-use premixed hybrid reagent simplifies reaction setup to only three components, reducing manual pipetting errors;

● All components are stored at uniformly at 2 ~ 8°C, eliminating repeated freeze-thaw cycles and minimizing cumbersome preparation steps;

● Wash-free beads design further streamlines operation and shortens the regular hybrid capture workflow to as little as 145 min.

Enhanced Compatibility for More Flexible Automation

● Supports 0.5-12 μg pooled library input and flexible hybrid times from 0.25-16 hr, balancing sample throughput with cost efficiency;

● Compatible with mainstream NGS platforms such as NovaSeq and DNBSEQ, with different adapter options to support multi-platform deployment;

● Simplifies automated workflow configuration and script development, enabling rapid automation deployment while significantly reducing instrument implementation and method validation costs.

Broad Probe Compatibility for Comprehensive Applications

● Conventional Probes: Deliver uniform and stable capture performance, supporting multiple variant analyses for routine targeted sequencing applications such as companion diagnostics and MRD monitoring;

● MSRE Probes: Enable simultaneous detection of DNA mutations and methylation, supporting dual-biomarker analysis for early cancer signal identification;

● Exhaustive Methylation Probes: Provide accurate quantification of methylation levels, supporting methylation biomarker discovery and validation as well as multi-cancer early screening product development.

workflow

Note: The hybrid capture of Conventional DNA Library is shown as an example. Regular refers to the μCaler Hybrid Capture Reagents v2, while HyperMix refers to the μCaler HyperMix Hybrid Capture Kit.

* Hybrid time is ﬂexible from 0.25-16 hr. 1 hr is recommended for Conventional DNA Library and 2 hr for Methylated DNA Library.

** Set the appropriate elution temperature according to the library type for use: 60℃ for Conventional DNA Library and 61℃ for Methylated DNA Library.

Performance

Ultra-large Pooled Library Input Capacity

Fig.1

Figure 1. Capture performance of μCaler HyperMix Hybrid Capture Kit across diﬀerent pooled library input amounts. Pre-libraries were prepared using the NadPrep DNA Library Preparation Module v2, coupled with the NadPrep Universal Stubby Adapter (UDI) Module. Hybrid capture was performed with pooled library input amounts of 0.5, 3, 6, and 12 μg using the M71 and the μCaler HyperMix Hybrid Capture Kit (for Illumina®) (1-hr hybridization). Sequencing was performed on NovaSeq 6000 (PE150). For each sample, 0.3 Gb data was used for subsequent analysis. A. Mappability and On-target rate; B. Target covered; C. Fold 80 base penalty; D. Average sequencing depth after deduplication.
Note: Samples were Human Genomic DNA standard (Promega, G1471), with an initial input amount of 50 ng.

Fully Compatible with Mainstream Sequencing Platforms

Fig.2

Figure 2. Capture performance of two μCaler hybrid capture reagents across diﬀerent sequencing platforms.Pre-libraries were prepared using the NadPrep DNA Library Preparation Module v2 coupled with either the NadPrep Universal Stubby Adapter (UDI) Module or the NadPrep Universal Adapter (MDI) Module (for MGI). 500 ng of each pre-library was performed to hybrid capture using the μCaler Hybrid Capture Reagents v2 (Regular) or the μCaler HyperMix Hybrid Capture Kit (for Illumina®/MGI) (HyperMix) with the M71. Sequencing was performed on the NovaSeq 6000 (PE150) and DNBSEQ-T7 (PE150) platforms. A. Mappability and On-target rate; B. Target covered; C. Fold 80 base penalty; D. Average sequencing depth after deduplication.

Consistent Detection Across Multiple Variant Types

Fig.3

Figure 3. Consistency between observed variant allele frequencies (VAFs) and the expected VAFs in reference standards using two μCaler hybrid capture reagents. 500 ng of each pre-library was performed to hybrid capture using Regular or HyperMix with the μCaler Custom Panel. Variant analysis was conducted using VarDict.

Note: Samples were PancancerLight 800 gDNA Reference Standard (GeneWell, GW-OGTM800), with an initial input amount of 50 ng.

Accurate Detection for Reliable DNA Methylation Analysis

Fig.4

Figure 4. Capture performance of two μCaler hybrid capture reagents using simulated samples with varying methylation levels. Pre-libraries were prepared from 50 ng simulated samples using the NadPrep Methyl Library Preparation Module v2 and NadPrep Methyl Stubby Adapter (UDI) Module v2 (with 10 nt index) after bisulfite conversion. 500 ng of each pre-library was performed to hybrid capture using Regular or HyperMix (2-hr hybridization) with the μCaler EMS Panel v1.0. A. Average coverage depth across all CpG sites. B. CpG methylation levels in target regions.

Note: The samples consist of 100% Methylated DNA (Zymo, D5014-2) as a positive control and 0% Methylated DNA (Zymo, D5014-1) as a negative control, mixed at different ratios to simulate methylation levels (0%, 10% and 50%).

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

μCaler HyperMix Hybrid Capture Kit (for Illumina®)

Color of Tube Cap	Component	Volume 1105302 (16 rxn)	Volume 1105301 (96 rxn)
Color of Tube Cap	Component	Volume 1105302 (16 rxn)	Volume 1105301 (96 rxn)
	μHybCap HyperMix (for Illumina®)	890 μL	3×1,770 µL
	μEnhancer	1,300 μL	8 mL
	Hyper Streptavidin Beads	160 μL	920 μL
/	Wash Buffer A Pro	5.8 mL	2×17.5 mL
/	Wash Buffer B	2.9 mL	20 mL

μCaler HyperMix Hybrid Capture Kit (for MGI)

Color of Tube Cap	Component	Volume 1105402 (16 rxn)	Volume 1105401 (96 rxn)
Color of Tube Cap	Component	Volume 1105402 (16 rxn)	Volume 1105401 (96 rxn)
	μHybCap HyperMix (for MGI)	890 μL	3×1,770 µL
	Hyper Streptavidin Beads	160 μL	920 μL
/	Wash Buffer A Pro	5.8 mL	2×17.5 mL
/	Wash Buffer B	2.9 mL	20 mL

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

What are the possible reasons for relatively high library yield after hybrid capture with μCaler? How to solve it?

Title: What are the possible reasons for relatively high library yield after hybrid capture with μCaler? How to solve it?

Possible Reason	Solution
Relatively high starting input	Quantification by Qubit™ 3.0 is recommended.
Relatively high amplification cycles	Explore the optimal amplification cycles according to the recommended protocol in the operating guideline.
The elution is not performed according to operational guideline, leading to serious off-target.	The elution is performed in strict accordance with the requirements of operating guideline.

What are the possible reasons for low captured-library yeild? How to solve it?

Title: What are the possible reasons for low captured-library yeild? How to solve it?

Possible Reason	Solution
Inaccurate starting input	In case of lower actual starting input, precise quantification by qPCR is recommended.
Reaction mixture is not completely transferred out when pipetting.	Transfer all reaction mixture from each step to a new tube when pipetting.
Some magnetic beads are discarded during hybridization and bead purification	Avoid too intense operation during the elution, and discard supernatant without losing any magnetic beads.

What are the possible reasons for low on-target rate after hybrid capture with μCaler? How to solve it?

Title: What are the possible reasons for low on-target rate after hybrid capture with μCaler? How to solve it?

Possible Reason	Solution
A lot of air bubbles are generated during elution	When using a pipette, it is necessary to gently pipette up and down to mix well to prevent the air from entering; If air bubbles are generated, first use the pipette to remove the air bubbles above the solution and then discard the supernatant.
PCR tube cap is not replaced	Replace the PCR tube and cap in strict accordance with the instruction for use.
Reagent is not fully mixed	Mix the reagents thoroughly before use according to the operation guideline and allow to warm up at the corresponding temperature.

Product	Catalog#
μCaler HyperMix Hybrid Capture Kit (for Illumina®), 16 rxn	1105302
μCaler HyperMix Hybrid Capture Kit (for Illumina®), 96 rxn	1105301
μCaler HyperMix Hybrid Capture Kit (for MGI), 16 rxn	1105402
μCaler HyperMix Hybrid Capture Kit (for MGI), 96 rxn	1105401