Double-Stranded RNA Sequencing (dsRNA-Seq)

CD Genomics utilizes cutting-edge double-stranded RNA (dsRNA) sequencing technology to identify and characterize RNA viruses, both plant and animal RNA viral infections without prior knowledge, enabling proactive containment of emerging infectious agents.

Overview

In the realm of viral infections, dsRNA plays a crucial role as it constitutes the viral genome in dsRNA viruses or is produced within host cells during viral replication. Recognizing the significance of dsRNAs, organisms across the board have developed the ability to detect and respond to these molecules, aiming to combat potential infections effectively.

Our dsRNA Sequencing service leverages high-throughput sequencing of total RNA isolated from infected individuals to identify and study the complete genomes of RNA viruses. This approach is particularly valuable when dealing with unknown pathogens causing diseases. By employing this method, we can also detect variants of known viruses or synthetic viral agents that may not be identifiable through traditional techniques such as PCR or serology-based assays.

At CD Genomics, we have implemented a rigorous viral enrichment and dsRNA purification process, which significantly reduces the presence of rRNA sequences. This meticulous purification step enhances the sensitivity of sequencing and improves our ability to reconstruct viral genomes accurately.

With our advanced dsRNA-Seq technology, we are capable of identifying and assembling the complete genomes of various RNA viruses. Our service covers a broad range of viral classifications, including negative-sense RNA viruses, positive-sense RNA viruses, dsRNA viruses, and bisense RNA viruses. This versatility allows us to effectively identify plant and animal RNA viral infections without relying on a priori knowledge, thus enabling a proactive approach in containing emerging infectious agents.

Features

High Compatibility Sensitivity and Accuracy Cutting-edge Algorithms One-Stop Solution
A wide range of viral families, including negative-sense RNA viruses, positive-sense RNA viruses, dsRNA viruses, and bisense RNA viruses. Stringent viral enrichment and dsRNA purification process, minimizing the presence of rRNA sequences, reducing background noise and improving the sensitivity of viral detection. This capability expands your ability to monitor viral evolution, track genetic changes, and enhance surveillance efforts, providing critical insights for public health and research purposes. One-stop solution from sample QC, library construction, to sequencing and data analysis, discovery of novel pathogens and the elucidation of their genetic makeup and potential implications.

Project Workflow

Sample Preparation

1. Sample Preparation

Library Preparation

2. Library Preparation

Sequencing

3. Sequencing

Data Analysis

4. Data Analysis

Bioinformatics Analysis Pipeline

1. Sequence Alignment: We align sequencing reads to reference genomes to identify known viruses and potential variants.

2. De novo Assembly: For novel or uncharacterized viruses, we perform de novo assembly to reconstruct their complete genomes.

3. Variant Calling: We identify genetic variations within viral genomes to study their diversity and evolution.

4. Functional Annotation: We annotate viral genomes to understand their potential functions and identify key genes or regions of interest.

5. Comparative Analysis: We compare identified viral genomes with known databases to gain insights into their relatedness and evolutionary relationships.

Case Studies

Sample Requirements

RNA sample (concentration ≥ 200 ng/uL, quantity ≥ 100 ug)

1.8 ≤ OD260/280 ≤ 2.2, OD260/230≥2.0, RIN ≥ 6.5, 28S:18S≥1.0.

Please make sure that the RNA is not degraded.

Sample Storage: RNA can be dissolved in ethanol or RNA-free ultra-pure water and stored at -80°C. RNA should avoid repeated freezing and thawing.

Shipping Method: When shipping RNA samples, the RNA sample is stored in a 1.5 mL Eppendorf tube, sealed with a sealing film. Shipments are generally recommended to contain 5-10 pounds of dry ice per 24 hours.

Deliverable: FastQ, BAM, coverage summary, QC report, custom bioinformatics analysis.

Reference:

  1. Izumi, Takuma, et al. "DsRNA sequencing for RNA virus surveillance using human clinical samples." Viruses 13.7 (2021): 1310.
* For Research Use Only. Not for use in diagnostic procedures.


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