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Nanodigmbio-μCaler Total Solution for SARS-CoV-2

Nanodigmbio-μCaler Total Solution for SARS-CoV-2

Nanodigmbio-μCaler Total Solution for SARS-CoV-2

Product catalog summary
Background
SARS-CoV-2, the virus responsible for COVID-19, is a positive-sense single-stranded RNA virus approximately 29.9 Kb in size. The genome is divided into two main parts: the 5'-proximal two-thirds encoding replicase polyproteins and the 3' third encoding structural and accessory proteins. The virus has a high mutation rate, leading to various variants such as Alpha, Beta, Gamma, Delta, and Omicron. Monitoring the virus's evolution and mutations is crucial for effective epidemic prevention. Targeted enrichment techniques, like liquid hybridization capture, enhance detection sensitivity and allow for the discovery of new variants.

SARS-CoV-2 Genome Structure
The genome includes ORF1a, ORF1b, and the Spike (S) protein, among others. A schematic diagram illustrates the genome structure, highlighting non-structural proteins and subunits.

Workflow
The document outlines a detailed workflow for preparing SARS-CoV-2 samples for sequencing. This includes steps like end repair, adapter ligation, and hybridization capture, using various modules such as the NadPrep Total RNA-To-DNA Module and the μCaler Hybrid Capture System.

Introduction
The NadPrep Total RNA-To-DNA Module is designed for NGS platforms, converting RNA to dsDNA. The NadPrep DNA Library Preparation Module prepares high-quality libraries for sequencing on MGI and Illumina platforms. The μCaler Hybrid Capture Reagents and NanoBlockers form a complete system for targeted enrichment and sequencing.

Features and Performance
The μCaler SARS-CoV-2 Panel offers a simplified process, completing the capture-library preparation in one day. It provides stable capture performance and excellent genome-wide coverage, as demonstrated in figures showing performance metrics and coverage depth.

Genotyping and Variation Map Analysis
Using the μCaler SARS-CoV-2 Panel, genotyping and variation analysis are performed on SARS-CoV-2 samples. The analysis uses the Wuhan-Hu-1/2019 reference genome for alignment, with results visualized in variation maps.

Ordering Information
The document provides a catalog of products related to the NadPrep and μCaler systems, including modules for RNA-to-DNA conversion, library preparation, and hybrid capture.

Statement
The products are for research use only and not for diagnostic procedures. Reproduction or translation of the document's content requires permission from Nanodigmbio.
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Catalog excerpts

Nanodigmbio-μCaler Total Solution for SARS-CoV-2-1

Severe acute respiratory syndrome coronavirus 2 (hereafter referred to as SARS-CoV-2) is the main pathogen causing novel co-ronavirus pneumonia (COVID-19); the SARS-CoV-2 is a positive-sense single-stranded RNA virus of ~ 29.9 Kb in size. The 5'-prox-imal two thirds of the SARS-CoV-2 genome encodes replicase polyproteins, and the 3' third of the genome encodes structural proteins and accessory proteins (the schematic diagram of genome structure is shown in Figure 1). A set of sgmRNAs (subgenomic mRNAs) that is consistent with the common leader sequence at the 5'-end and the 3'-end sequence of the virus genome sequence were finally generated using a discontinuous replication and transcription mechanism. Therefore, SARS-CoV-2 has strong mutation ability and is more likely to mutate compared with DNA viruses. Up to now, SARS-CoV-2 has evolved into Alpha, Beta, Gamma, Delta as well as Omicron variants. As the number of infected individuals increases, SARS-CoV-2 has evolved into many different subvariants. Although the current mainstream Omicron variants have very low pathogenicity and fatality rates in China, it does not mean that SARS-CoV-2 will not evolve into more virulent variants in the future. Therefore, determining the genomewide sequence of SARS-CoV-2, typing and subtyping the virus, exploring the evolutionary spectrum of the virus, and closely monitoring its variation are of great significance for precise epidemic prevention. In view of the low virus content of clinical samples, targeted enrichment can greatly improve the detection sensitivity, and common detection methods include long-fragment PCR amplification technique and liquid hybridization capture technique. Compared with amplicon, liquid hybridization capture technique has higher fault tolerance without requiring emergence of new variants before updating primer design in amplicon sequencing, and allows the discovery of new variants through long-term tracking with superior coverage uniformity. Therefore, Nanodigmbio developed the |iCaler SARS-CoV-2 Panel based on its proprietary |iCaler Hybrid Capture System, so as to provide a simple and rapid SARS-CoV-2 whole-genome sequencing solution with excellent coverage homogeneity. Figure 1. Schematic diagram of SARS-CoV-2 genome structure.

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Nanodigmbio-μCaler Total Solution for SARS-CoV-2-2

* The NadPrep Universal Stubby Adapter (UDI) is shown in the schematic diagram as an example.

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Nanodigmbio-μCaler Total Solution for SARS-CoV-2-3

NadPrep Total RNA-To-DNA Module is a reverse transcription module developed for NGS platforms. This module offers an efficient solution for the generation of dsDNA fragments from 10-200 ng of human total RNA from cells, tissues and FFPE samples. The dsDNA output can couple with the NadPrep DNA Library Preparation Kit-series. NadPrep DNA Library Preparation Module is designed for the preparation of high-quality libraries from double-stranded DNA (dsDNA) on MGI and Illumina® platforms. This A-T ligation based kits offers a stable and efficient library preparation solution for a variety of applications....

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Nanodigmbio-μCaler Total Solution for SARS-CoV-2-4

Figure 2. Libraries were prepared with RNA samples using the NadPrep Total RNA-To-DNA Module coupled with the NadPrep DNA Library Preparation Kit (for Illumina®), and a complete hybrid capture solution was developed using the |jCaler SARS-CoV-2 Panel coupled with the |_iCaler Hybrid Capture Reagents and |_iCaler NanoBlockers (for Illumina®), with sequencing mode of MiniSeq SE 100. The sequencing data were analyzed using IRMA. A. Capture performance of the |jCaler SARS-CoV-2 Panel on cell-passaged SARS-CoV-2; B. The genome-wide coverage of SARS-CoV-2. Figure 3. Capture performance and genome-wide...

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Nanodigmbio-μCaler Total Solution for SARS-CoV-2-5

Ordering Information Type RNA-To-DNA Lib Prep Module NadPrep DNA Library Preparation Kit (for Illumina®) G24 NadPrep Total RNA-To-DNA Module, 96 rxn NadPrep DNA Library Preparation Kit (for Illumina®) E96 NadPrep Universal Stubby Adapter (UDI) Module Set A1, 24 rxn NadPrep Universal Stubby Adapter (UDI) Module Set B2, 96 rxn NadPrep Universal Stubby Adapter (UDI) Module Set E1, 96 rxn Module NadPrep Universal Stubby Adapter (UDI) Module Set E2, 96 rxn NadPrep Universal Stubby Adapter (UDI) Module Set E3, 96 rxn NadPrep Universal Stubby Adapter (UDI) Module Set E4, 96 rxn NadPrep Universal Stubby...

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