NadPrep Methyl Library Preparation Kit

Fig 1.Library preparation performance of different types of samples and different conversion methods.A. Library yield of different types of samples，B. Library yield and C. λ DNA C conversion rate of of simulated samples with different methylation levels indicate stable performance and differences in conversion step treated by enzymatic (EM) and bisulfite (BS), respectively. The libraries were prepared by using NadPrep Methyl Library Preparation Module coupled with NadPrep Methyl Adapter (SI/MDI) Module (for MGI).
Note：The human 100% methylated DNA standard (Zymo, D5014-2) positive control and 0% methylated DNA standard (Zymo, D5014-1) negative control were mixed in different proportions to simulate different methylation levels (0%, 10%, 50% and 100%).

Fig 2.Library preparation performance of different conversion methods. The libraries were prepared by using NadPrep Methylation Library Preparation Module coupled with NadPrep Methyl Stubby Adapter (UDI) Module (for Illumina®) (Stubby) and NadPrep Methyl UDI Adapter Module (for Illumina®) (Full) in conversion step treated by enzymatic (EM) and bisulfite (BS), respectively. A. Library yield of two kinds of adapters in different conversion step. B. Library A yield of two kinds of stubby adapters in conversion step treated by EM. C. λ DNA C conversion rate in different conversion step.
Note：Samples are simulated DNA samples by using human 100% methylated DNA standard (zymo, D5014-2) positive control and 0% methylated DNA T standard (zymo, D5014-1) negative control to mimic different methylation levels (0%, 50% and 100%). Input amount: 50 ng. The adapter of B & C are Stubby.

Fig 3. Capture performance of libraries treated by different conversion methods.A. Mappability and On-target rate, B. Target covered, C. Coverage uniformity and consistency, and D. GC bias show satisfying performance treated by EM and BS, respectively.Note: Samples are 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%, 50% and 100%).

Fig 4. Capture performance of libraries treated by different conversion methods. The libraries were prepared by using NadPrep Methylation Library Preparation Module coupled with NadPrep Methyl Stubby Adapter (UDI) Module (for Illumina®). The capture was applied by a methylation demo panel (60 Kb), then sequenced on Novaseq 6000, PE 150. The A. Mappability and On-target rate, B. Target covered, C. Coverage uniformity and consistency, and D. GC bias show satisfying performance treated by EM and BS, respectively. 

Fig 5. NadPrep^® Performance of different methylation level samples treated by EM and BS solutions. The libraries were prepared by using NadPrep Methylation Library Preparation Module coupled with NadPrep Methyl Adapter (MDI) Module (for MGI). The capture was applied by a methylation demo panel (60 Kb), then sequenced on MGISEQ-2000, PE100. A. The actual detection results of samples with different methylation levels and B. The CpG methylation level of samples treated by EM and BS conversion solution are shown, respectively.Note: Samples are 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%, 50% and 100%). 

Fig 6. Performance of different methylation level samples treated by EM and BS solutions.  The libraries were prepared by using NadPrep Methylation Library Preparation Module coupled with NadPrep Methyl Stubby Adapter (UDI) Module (for Illumina®). The capture was applied by a methylation demo panel (60 Kb), then sequenced on Novaseq 6000, PE 150. A. The CpG methylation level of samples treated by EM conversion solution; B. The CpG methylation level of samples treated by BS conversion solution.