NadPrep RNA & DNA Library Co-Preparation Module

Figure 1. NadPrep RNA & DNA Library Co-Preparation Module for library co-preparation with mixed RNA & DNA samples in varying initial ratios. A. Library yield (MDI); B. Library yield (UDI). Utilize NadPrep RNA & DNA Library Co-Preparation Module in conjunction with NadPrep Universal Adapter (MDI) Module (for MGI) and NadPrep Universal Stubby Adapter (UDI) Module for library co-preparation. 
Note: The RNA samples are from K562 cell lines; the DNA samples are Human Genomic DNA Standards (Promega, G1521).

Figure 2. Detection of tNGS Total Solution with NadPrep RNA & DNA co-preparation on dual sequencing platforms. A. Mappability; B. Reads distribution of Coronavirus; C. Reads distribution of Candida albicans; D. Reads distribution of rRNA. Pre-libraries preparation were performed using NadPrep RNA & DNA Library Co-Preparation Module. Hybridization capture were completed with NadPrep ES Hybrid Capture Reagents and NEX-t Panel v1.0, followed by sequencing on DNBSEQ-T7, PE150 and Illumina NovaSeq 6000, PE150. For each sample, 0.5 Gb was used for data analysis.
Note: The scale on the X-axis represents the proportional initial input amounts of the host-mimicking mixed nucleic acid samples to the input amounts of the pathogenic mixed nucleic acid samples. The host-mimicking mixed nucleic acid samples consist of Human Brain Total RNA Standard (Clontech, 636530) and Human Genomic DNA Standard (Promega, G1521) mixed at a 1:1 ratio. The pathogenic mixed nucleic acid samples consist of the Reference material for SARS-CoV-2 Omicron Variant Genomic RNA (National Institute of Metrology, NIM-RM 5225; 1 pg equals 50 copies) and Candida albicans standard (ATCC, 10231; 1 pg equals 6000 copies) mixed at a 1:1 ratio.

Figure 3. Distribution of detected reads in simulated pathogenic microbial samples using tNGS and mNGS based on the NadPrep RNA & DNA Library CoPreparation Module. A. Reads distribution of Coronavirus; B. Reads distribution of Candida albicans.
Note: Sequencing platform: Illumina NovaSeq 6000, PE150.

Figure 4. Capture performance of mixed RNA samples in varying copy numbers with co-preparation and RNA-preparation. A. Reads distribution; B. Enrichment fold.
Note: The X-axis represents the copy number of the Reference material for SARS-CoV-2 Omicron Variant Genomic RNA. The samples are derived from a mixture of Human Brain Total RNA Standard (Clontech, 636530) and Omicron virus samples in varying copy numbers.

Figure 5. Captured coverage depth after duplication removal for samples with different microbial contents with co-preparation and DNA-preparation.The simulated microbial community samples were prepared using the NadPrep RNA & DNA Library Co-Preparation Module (Co-Prep) and NadPrep EZ DNA Library Preparation Module v2 (DNA-Prep), in conjunction with NadPrep Universal Stubby Adapter (UDI) Adapter for library preparation, followed by hybrid capture using NadPrep ES Hybrid Capture Reagents and the NEX-t Panel v1.0.
Note: Simulated microbial community samples of 0.001% - 1% MSA-1003 were created by diluting a mixture of 20 strains of genomic material (ATCC, MSA-1003) using the human genomic DNA standard (Promega, G1471) at various ratios. Sequencing platform: Illumina NovaSeq 6000, PE150.

Figure 6. Uniform coverage of regions with various GC content using NadPrep RNA & DNA Library Co-Preparation Module. A. Staphylococcus aureus, B. Bacillus subtilis, and C. Lactobacillus fermentum with different GC content.

Figure 7. Detection performance of tNGS Total Solution for clinical samples with NadPrep RNA & DNA co-preparation. A. Library yield; B. Reads ratio of Rhinovirus A and Rhinovirus C. 
Note: The samples were derived from 50 ng of clinical rhinovirus nucleic acid samples. Sequencing platform: Illumina NovaSeq 6000, PE150