NadPrep ES Hybrid Capture Reagents v2

Catalog Number:

#1005422
#1005421

NadPrep ES Hybrid Capture Reagents v2 is an upgraded rapid hybridization and wash reagent kit optimized for targeted capture using 120 nt NAD Probes/Panels. Together with the NadPrep NanoBlockers series and 120 nt NAD Probes/Panels, it forms a complete liquid-phase hybrid capture system. This system supports ﬂexible optimization of capture performance across panels of diﬀerent sizes by adjusting hybridization time, and enables ﬂexible selection of hybridization durations from 1-16 hours as well as multiplex library pooling from 1-12 plex.

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Feature

● Streamlined System: Only 1 hybridization premix + 1 wash buﬀer are required, signiﬁcantly reducing reagent preparation steps, minimizing operational errors, and improving automation compatibility.

● Uniﬁed Conditions: Only 2 incubation temperatures are involved, avoiding frequent temperature adjustments and making the workﬂow smoother and easier to scale.

● Optimized Workﬂow: A workﬂow design of 2 beads washes + 3 heated washes + 1 ethanol wash, greatly improving exper-imental eﬃciency while maintaining reliable performance.

● Flexible Compatibility: Supports 1-16 hr hybridization time and 1-12-plex library pooling, allowing ﬂexible adjustment according to experimental needs to achieve optimal capture performance.

● Stable and Reproducible: A highly standardized workﬂow reduces dependence on operator experience, enabling diﬀerent users to obtain stable and consistent capture results.

● Comprehensive Improvement: Eﬀectively improves captured library yield, target region coverage, and coverage unifor-mity, while reducing GC bias, minimizing rework, and lowering sequencing costs.

Workflow-01

Note：Nad-Traditional refers to NadPrep Hybrid Capture Reagents; NadPrep ES v1 and v2 refer to NadPrep ES Hybrid Capture Reagents v1 and v2, respectively.

Performance

Uncompromising Multi-dimensional Capture Performance: ES v2 1_hr Rapid Hybrid vs. ES v1 16_hr Overnight Hybrid

Fig 1. Capture performance of NadPrep ES Hybrid Capture Reagents v2 under diﬀerent hybridization times. Pre-libraries were prepared using the NadPrep EZ DNA Library Preparation Kit v3. 500 ng of each pre-library was used for hybridization capture with panels of diﬀerent sizes using NadPrep ES Hybrid Capture Reagents v1 (NadPrep v1) and v2 (NadPrep v2). Sequencing was performed on NovaSeq 6000 (PE150). For each sample, appropriate amount of data was randomly selected for subsequent analysis. On-target rate was calculated by the number of reads. A. Mappability and On-target rate; B. Target covered; C. Fold 80 base penalty; D. Captured-library yield.
Note：Samples were Human Genomic DNA Standard (Promega, G1471) and Cell Line gDNA Standard (Coriell, NA12878).

Excellent Capture Stability Across Panels of Diﬀerent Sizes

figure.2

Fig 2. Capture performance of NadPrep ES Hybrid Capture Reagents v2 across panels of diﬀerent sizes. Pre-libraries were prepared using NadPrep EZ DNA Library Preparation Kit v3. 500 ng of each pre-library was used for hybrid capture using panels of diﬀerent sizes, combining with NadPrep v2 and Vendor A (1-hr hybridization). Sequencing was performed on NovaSeq 6000 (PE150). A. Mappability and On-target rate; B. Target covered; C. Fold 80 base penalty; D. Captured-li-brary yield.

Support for Multiplex Library Hybridization Across Diﬀerent Sequencing Platforms

figure.3

Fig 3. Capture performance of NadPrep ES Hybrid Capture Reagents v2 across diﬀerent multiplex hybridization conditions. Pre-libraries were prepared using NadPrep EZ DNA Library Preparation Kit v3. Hybridization was performed using either 500 ng per pre-library (1-plex) or 6,000 ng total input for 12 pre-libraries (12-plex) across panels of diﬀerent sizes, combining with NadPrep v2. Sequencing was performed on NovaSeq 6000 (PE150) and DNBSEQ (PE150) respectively. A. Mappability and On-target rate; B. Target covered; C. Fold 80 base penalty; D. Captured-library yield; E. GC bias (2.4 Mb, 1-hr hybridization); F. GC bias (41.7 Mb, 1-hr hybridization).

Robust Detection of Multiple Variant Types

figure.4

Fig 4. Consistency between observed variant allele frequencies (VAFs) and the expected VAFs in reference standards using NadPrep ES Hybrid Capture Reagents v2. Pre-libraries were prepared using NadPrep EZ DNA Library Preparation Kit v3. 500 ng of each pre-library was used for hybrid capture using LungCancer Panel v1.0, combining with NadPrep v2 and v1, as well as Vendor A (1-hr hybridization). Sequencing was performed on NovaSeq 6000 (PE150), and variant analysis was conducted using Vardict.
Note：Samples were PancancerLight 800 gDNA Reference Standard (GeneWell, GW-OGTM800), with an input amount of 50 ng.

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

Package	Color Of Tube Cap	Component	Volume 1005422 (16 rxn)	Volume 1005421 (96 rxn)
Package	Color Of Tube Cap	Component	Volume 1005422 (16 rxn)	Volume 1005421 (96 rxn)
Box 1		ES Hyb Mix	290 μL	2×1,750 μL
		Human Cot DNA	110 μL	650 μL
		Sealing Oil	250 μL	1,500 μL
Box 2	or /	Streptavidin Beads II	950 μL	5.7 mL
Box 2	/	ES Wash Buffer II	7.5 mL	3×30 mL

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

Is NadPrep ES Hybrid Capture Reagents v2 compatible with the bead-based concentration method?

NadPrep ES Hybrid Capture Reagents v2 is compatible with the bead-based concentration (BC) method. Compared with the vacuum concentration (VC) method, BC method is simpler to operate, faster, and more suitable for automated platforms (see the Appendix of the user manual for details). It should be noted that due to differences in purification recovery efficiency, the captured library yield may be slightly lower when using BC method compared with VC method.

Product

Catalog

NadPrep ES Hybrid Capture Reagents v2, 16 rxn

1005422

NadPrep ES Hybrid Capture Reagents v2, 96 rxn

1005421

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