NadPrep EZ RNA & DNA Library Co-Preparation Module

Figure 1. Library preparation performance of the NadPrep EZ RNA & DNA Library Co-Preparation Kit for mixed RNA and DNA samples across different input amounts. The libraries were prepared using the NadPrep EZ RNA & DNA Library Co-Preparation Module coupled with the NadPrep Universal Stubby Adapter (UDI) Module (NovaSeq) and the NadPrep Universal Adapter (MDI) Module (for MGI) (DNBSEQ). A. Library yield; B. Size distribution of the libraries (NovaSeq).

Note: RNA samples were extracted from the Jurkat cell line. DNA samples were Human Genomic DNA standards (Promega, G1521).

Table 1. Evaluation of microbial residues in the NadPrep EZ RNA & DNA Library Co-Preparation Kit using an in-house background microbial database.

Metagenomic libraries were prepared from host-mimicking mixed nucleic acid samples using the NadPrep EZ RNA & DNA Library Co-Preparation Kit. The host-mimicking mixed nucleic acid samples consist of Human Brain Total RNA (Clontech, 636530) and Human Genomic DNA standard (Promega, G1521) mixed at 50 ng:50 ng. Sequencing was performed on NovaSeq 6000 with PE150. The average raw data was 5 M reads pair per sample.

Table 2. Comparison of pathogen detection performance between two RNA & DNA library co-preparation kits for reference standards across different concentration gradients.

A total of 400 μL of reference standards at different concentration gradients were used for DNA extraction. Pre-libraries were prepared using the NadPrep RNA & DNA Library Co-Preparation Kit (Co-Prep) and NadPrep EZ RNA & DNA Library Co-Preparation Kit (EZ Co-Prep), respectively. 50% of the pre-libraries were performed to hybridization capture using the μCaler HyperMix Hybrid Capture Kit (for Illumina®) and the μCaler RIA Panel v1.0. Sequencing was performed on NovaSeq 6000 with PE150. 0.5 Gb of raw data per sample was used for analysis.

Note: ◉ Detected; ◉ Not Detected. Initial input templates were used at the maximum nucleic acid volume recommended in the corresponding user manual.

Figure 3. Library preparation performance and pathogen detection capability of two RNA & DNA library co-preparation kits using oropharyngeal swab samples from patients with respiratory mixed infections. RNA and DNA mixed nucleic acids were extracted from 400 μL of swab preservation medium. Pre-libraries were prepared using the two kits, respectively. 50% of the pre-libraries were performed to hybridization capture using the μCaler HyperMix Hybrid Capture Kit (for Illumina®) and μCaler RIA Panel v1.0. A. Pre-library yield; B. RPM of detected pathogens.

Table 3. Library preparation performance and HIV-1 whole-genome sequencing capability of the NadPrep EZ RNA & DNA Library Co-Preparation Kit using blood samples from HIV-infected individuals.

RNA was extracted from appropriate amount of blood sample. Pre-libraries were prepared using the NadPrep EZ RNA & DNA Library Co-Preparation Kit. 50% of the pre-libraries were performed to hybridization capture using the μCaler HyperMix Hybrid Capture Kit (for Illumina®) and μCaler HIV-1 Panel. Analysis support: EGA HIV-1 Subtyping Tool - Version 3.0 (dbpartners.stanford.edu:8080/RegaSubtyping/stanford-hiv/typingtool/)