NadPrep Methyl Library Preparation Kit v2

Catalog Number:

#1002511
#1002512

NadPrep Methyl Library Preparation Kit v2 is designed for preparation of high-quality methylation sequencing (Methyl-Seq) libraries from double-stranded DNA (dsDNA) on both Illumina and MGI platforms. This kit is suitable for preparing Methyl-Seq libraries from 1-500 ng DNA, supporting whole-genome bisulfite sequencing (WGBS) as well as hybrid capture-based targeted methylation sequencing with effective enrichment of targeted libraries.

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Feature

Supports input amounts ranging from 1-500 ng across different DNA sample types, enabling better sample utilization.
Compatible with various conversion methods and sequencing platforms, offering flexibility for diverse applications.
Enables WGBS library preparation in as fast as 4.5 hr with bisulfite conversion.
Significantly improves effective sequencing depth and data utilization.

Workflow

workflow

Performance

Efficient Library Yield

Figure 1. NadPrep Methyl Library Preparation Kit v2 achieves stable and efficient library yield across various applications. A. gDNA samples of varying input amounts processed with NadPrep Methyl Library Preparation Module v1 and v2 after BS conversion; B. gDNA samples of varying input amounts with EM; C. Library preparation for gDNA, FFPE DNA, and cfDNA with BS; D. Simulated samples with varying methylation levels with BS.
Note: The samples in figure 1.D were simulated DNA by using human 100% Methylated DNA standard (Zymo, D5014-2) positive control and 0% Methylated DNA standard (Zymo, D5014-1) negative control to mimic different methylation levels (0%, 10%, and 50%), with an initial input amount of 50 ng. BS: Bisulfite conversion; EM: Enzymatic conversion.

Stable Conversion Efficiency

Figure 2. Conversion efficiency across multiple types of samples at varying input amounts using different methods. Library preparation was performed using NadPrep Methyl Library Preparation Module v2 coupled with NadPrep Methyl Stubby Adapter (UDI) Module (with 10 nt Index). A. Conversion efficiency of gDNA across varying input amounts processed with different conversion methods; B. Conversion efficiency of multiple sample types processed with BS. The sequencing mode was Illumina Novaseq 6000 platform, PE150.

Improved Data Utilization

Figure 3. Capture performance of NadPrep Methyl Library Preparation Kits v1 and v2. A. Mappability & On-target rate; B. Target covered; C. Average sequencing depth after deduplication; D. GC bias. Pre-libraries were prepared from gDNA samples with various input amounts using NadPrep Methyl Library Preparation Module v1 & v2 coupled with NadPrep Methyl Stubby Adapter (UDI) Module (with 10 nt Index) and treated by BS. Hybrid capture was performed with NadPrep Hybrid Capture Reagents and Methyl Demo Panel (60 Kb). 0.3 Gb were randomly selected for data analysis.

Accurate Quantification of Methylation Levels

Figure 4. NadPrep Methyl Library Preparation Kit v2 precisely quantifies methylation levels. A. Detected methylation levels across simulated samples with varying methylation levels; B. CpG methylation levels in target regions. Pre-libraries were prepared from 50 ng simulated samples with different methylation levels using NadPrep Methyl Library Preparation Module v2 coupled with NadPrep Methyl Stubby Adapter (UDI) Module (with 10 nt Index) and treated by BS. Hybrid capture was conducted with μCaler Hybrid Capture Reagents v2 and μCaler Methyl Demo Panel (5 Kb).

Flexible Compatibility with Ultra-Sensitive μCaler Hybrid Capture System

Figure 5. NadPrep Methyl Library Preparation Kit v2 demonstrates excellent capture performance with low-input gDNA samples. A. Library yield; B. Mappability & On-target rate; C. Target covered; D. Average sequencing depth after deduplication. Pre-libraries were prepared using NadPrep Methyl Library Preparation Module v2 coupled with NadPrep Methyl Stubby Adapter (UDI) Module (with 10 nt Index) and treated by EM. Hybrid capture was performed with μCaler Hybrid Capture Reagents v2 and μCaler EMS Panel (20 Kb).

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

Lib Prep Module

#Package	Cap Color	Component	Volume 1002511 (24 rxn)	Volume 1002512 (96 rxn)
Box1		EndRepair & A-Tailing Buffer Pro	180 µL	690 µL
		End Repair & A-Tailing Enzyme Pro	120 µL	460 µL
		Ligation Buffer	750 µL	2×1,500 µL
		DNA Ligase	65 µL	250 µL
		2X HiFi-Methyl PCR Master Pro Mix	720 µL	2×1,600 µL
	/	Nuclease Free Water	2.5 mL	8 mL
		TE Solution	1 mL	4 mL
Box2	/	NadPrep SP Beads	4 mL	15 mL

Methyl Adapter (SI) Module (for MGI)

Cap Color	Component	Volume 1003631 (24 rxn)	Volume 1003632 (96 rxn)	Volume 1003633 (96 rxn)
	NadPrep M-Index (SI) Primer Mix #	12×15 µL	24×25 µL	24×25 µL
	NadPrep Methyl M-Adapter	60 µL	230 µL	230 µL

Methyl Adapter (MDI) Module (for MGI)

Cap Color	Component	Volume 1003740 (24 rxn)	Volume 1003741 (96 rxn)	Volume 1003742 (96 rxn)	Volume 1003743 (96 rxn）	Volume 1003744 (96 rxn）
	NadPrep M-Index (MDI) Primer Mix #	12×15 µL	24×25 µL	24×25 µL	24×25 µL	24×25 µL
	NadPrep Methyl M-Adapter	60 µL	230 µL	230 µL	230 µL	230 µL

Methyl Stubby Adapter (UDI) Module (for Illumina^®)

Cap Color	Component	Volume 1003371A (24 rxn)	Volume 1003381A (96 rxn)	Volume 1003382A (96 rxn)	Volume 1003391A (96 rxn)
	NadPrep UDI-Index Primer Mix II #	12×15 µL	24×25 µL	24×25 µL	96×8 µL
	NadPrep Methyl Stubby Adapter II	60 µL	230 µL	230 µL	260 µL
	NadPrep Amplification Primer Mix II	20 µL	75 µL	75 µL	75 µL

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

Type	Product	Detail	Catalog
Lib Prep Module	NadPrep Methyl Library Preparation Module v2, 24 rxn	24 rxn	1002511
Lib Prep Module	NadPrep Methyl Library Preparation Module v2, 96 rxn	96 rxn	1002512
BS Conversion Module	NadPrep DNA Methyl Bisulfite Conversion Module, 24 rxn	24 rxn	1002701
BS Conversion Module	NadPrep DNA Methyl Bisulfite Conversion Module, 96 rxn	96 rxn	1002702
Methyl Adapter (SI) Module (for MGI)	NadPrep Methyl Adapter (SI) Module Set A1 (for MGI), 24 rxn	SI 1-12	1003631
	NadPrep Methyl Adapter (SI) Module Set B1 (for MGI), 96 rxn	SI 1-24	1003632
	NadPrep Methyl Adapter (SI) Module Set B2 (for MGI), 96 rxn	SI 25-48	1003633
Methyl Adapter (MDI) Module (for MGI)	NadPrep Methyl Adapter (MDI) Module Set A1 (for MGI), 24 rxn	MDI 1-12	1003740
	NadPrep Methyl Adapter (MDI) Module Set B1 (for MGI), 96 rxn	MDI 1-24	1003741
	NadPrep Methyl Adapter (MDI) Module Set B2 (for MGI), 96 rxn	MDI 25-48	1003742
	NadPrep Methyl Adapter (MDI) Module Set B3 (for MGI), 96 rxn	MDI 49-72	1003743
	NadPrep Methyl Adapter (MDI) Module Set B4 (for MGI), 96 rxn	MDI 73-96	1003744
Methyl Stubby Adapter (UDI) Module (for Illumina^®)	NadPrep Methyl Stubby Adapter (UDI) Module v2 Set A1 (for Illumina^®), 24 rxn	UDI 1-12	1003371A
	NadPrep Methyl Stubby Adapter (UDI) Module v2 Set B1 (for Illumina^®), 96 rxn	UDI 1-24	1003381A
	NadPrep Methyl Stubby Adapter (UDI) Module v2 Set B2 (for Illumina^®), 96 rxn	UDI 25-48	1003382A
	NadPrep Methyl Stubby Adapter (UDI) Module v2 Set E1 (for Illumina^®), 96 rxn	UDI 1-96	1003391A