NadPrep EZ DNA Library Preparation Kit v3

Figure 1. Library yield of NadPrep EZ DNA Library Preparation Kit v3 and v2 for gDNA samples with different input amounts. The libraries were prepared by using NadPrep EZ DNA Library Preparation Module v3 (NadPrep Enfrag v3) and v2 (NadPrep Enfrag v2) coupled with A. NadPrep Universal Stubby Adapter (UDI) Module and B. NadPrep Universal Adapter (MDI) Module (for MGI) respectively, with 20 min of enzymatic digestion and recommended PCR cycles.
Note: The samples are human genome DNA standard (Promega, G1471).

Figure 2. Library yield of NadPrep EZ DNA Library Preparation Kit v3 and v2 for different types of samples.
Note: The grading standards by electrophoresis are as follows: gDNA: one bright and non-trailing stripe with about 15 kb in size. FFPE B: one indistinct stripe with about 15 kb in size, with medium diffusion. FFPE C: multiple stripes ranging from 200 bp to 2,500 bp, with severe diffusion. FFPE D: multiple stripes ranging from 250 bp to 1,000 bp, with severe diffusion.

Figure 3. The abnormal reads ratio of libraries prepared with different types of samples by using different library preparation kits. The libraries were prepared with gDNA and different grades of FFPE DNA samples (50 ng each) with different library preparation kits including NadPrep Enfrag v3, NadPrep Enfrag v2 and Vendor A*. The multi-hybridization reaction was initiated with 500 ng of each library with the same library preparation scheme. The capture was performed by using NanOnco Plus Panel v3.0 and NadPrep Hybrid Capture Reagents, with calculation of total reads. Sequencing was performed on (A) NovaSeq 6000, PE150 and (B) DNBSEQ-T7, PE150.
Note: Artifact rate: the ratio of abnormal sequence introduced by enzymatic digestion by-products. The initial input amounts of gDNA and FFPE DNA were 50 ng and 100 ng respectively.

Figure 5. Variant statistics from the capture data of Onco Wildtype gDNA Reference Standard using different library preparation kits. The capture was performed using NanOnco Plus Panel v3.0 and NadPrep Hybrid Capture Reagents. For each sample, 10 M read pairs were randomly selected and deduplicated for data analysis. Variants were called with a background noise frequency ranging from 0 to 0.005 (at least 3 reads per site).
Note: The samples are Onco Wildtype gDNA Reference Standard (GeneWell, GW-OGTM005), with an initial input of 50 ng.

Figure 6. The capture performance of pre-libraries prepared by NadPrep EZ DNA Library Preparation Kit v3. For each sample, 2 M read pairs were randomly selected for data analysis, with calculation of total reads. A. Mappability and On-target rate; B. Target covered; C. Fold 80 base penalty; D. Chimera rate (chimeric reads/total reads).