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NadPrep DNA Methyl Bisulfite Conversion Module

Catalog Number:

#1002701
#1002702

NadPrep DNA Methyl Bisulfite Conversion Module is a methylation conversion reagent developed for DNA samples based on the principle of bisulfite conversion. The conversion module, based on a magnetic bead purification protocol, efficiently conv- erts unmethylated cytosine to uracil within 2 hr, leaving methylated cytosine unchanged, and it can be compatible with autom- ated workstations. It can be seamlessly integrated with Nanodigmbio's NadPrep double-strand methylation library preparation scheme and NadPrep single-strand methylation library preparation scheme, allowing flexible selection among various types of starting samples.

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Bisulfite conversion workflow

*The above schematic diagram of adapter-ligated product is taking NadPrep Methyl Stubby Adapter (UDI) as an example.

Feature

Compatible with multiple types of samples and supports a wide range of input amounts
Efficient and stable cytosine conversion rate
Excellent detection sensitivity of 5mC and 5hmC
Convenient operation is conducive to automation implementation

Performance

Library yield and conversion efficiency

Fig 1. Library yield and conversion efficiency of samples with different input amounts.Human Genomic DNA standards (Promega, G1471) were used with NadPrep Methyl Library Preparation Module in combination with NadPrep Methyl Stubby Adapter (UDI) Module (with 10 nt Index) and NadPrep DNA Methyl Bisulfite Conversion Module for library preparation, Sequencing were performed on Illumina Novaseq 6000, PE150.A. Library yield；B. λ DNA C conversion rate.

Capture Performance

Fig 2. Capture performance of samples with different methylation levels. 50 ng simulated samples with different methylation levels were prepared using NadPrep Methyl Library Preparation Module in combination with NadPrep Methyl Stubby Adapter (UDI) Module (with 10 nt Index) after bisulfite conversion. The pre-libraries were capture with a methylation demo panel (60 Kb). Sequencing was performed on Illumina Novaseq 6000, PE150, 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. Average sequencing depth after deduplication.

Note：The samples consisted 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 different methylation levels (0%, 50%, and 100%). Vendor Q* and Vendor Z* refer to different bisulfite conversion modules provided by Vendor Q* and Vendor Z*, respectively.

Methylation detection

Fig 3. Targeted sequencing performance of samples with different methylation levels. A. Comparison between theoretical methylation levels and detected methylation levels; B. CpG methylation levels in target regions.

Package	Color of Cap Tube	Component	Volume 1002701 (24 rxn)	Volume 1002702 (96 rxn)
Package	Color of Cap Tube	Component	Volume 1002701 (24 rxn)	Volume 1002702 (96 rxn)
Box 1	/	Bisulfite Diluent	4.5 mL	9 mL
		Bisulfite Mix	4×1 tube	8×1 tube
		Desulf Buffer	1.8 mL	7.5 mL
Box 2	/	Nuclease Free Water	2.5 mL	8 mL
	/	Bisulfite Beads	4.8 mL	3×6.2 mL
		DNA Protect Buffer	100 μL	380 μL

Type	Product	Scale	Catalog
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