EasySeq™ RC-PCR SARS CoV-2 (novel coronavirus) Whole Genome Sequencing

Generate Whole Genome Sequencing data from cDNA, derived from Covid-19 positive tested patients.

Introduction

Info

The analysis and study of viral genomes is an essential epidemiological requirement to track the evolution of a virus, the origin of strains and their transmission during any epidemic or pandemic. Targeted Next Generation Sequencing (NGS) is a valuable tool for both detection and classification of viral genomes. The 2019 outbreak of the novel Coronavirus strain (SARS-CoV-2) is obviously unprecedented with over 26 million confirmed cases of the disease associated with SARS-CoV-2 (COVID-19) and sadly approaching 1 million of lost lives in a relatively short period of time. Therefore, the ability for public health professionals and researchers to monitor situations in the most efficient and streamlined manner possible is essential. As is the reassurance that testing methods are secure, and results are accurate.

RC-PCR, the next revolution in NGS Library Prep

– More efficient ­library prep: Up to 80% less hands-on time
– Combined Amplification and Indexing with RC-PCR
– Less Handling, Less Risk, Greater Sample Safety
– Easily adapted for Automation
– Single Click Analysis with virSEAK (JSI Medical Systems)

With the NimaGen EasySeq™ RC-PCR Library Preparation method the implementation of an NGS driven viral surveillance strategy could not be simpler or safer. Utilising patented RC-PCR technology, sequence ready libraries are generated from cDNA covering the entire 29.9Kb SARS-CoV-2 Genome with minimal hands-on time and just a single PCR for both amplification and indexing. Tiled RC-PCR primers and probes are provided in tubes, as well as 2 breakable 96 well plates which include UDI Indexes. Just add cDNA and the RC-PCR mastermix provided in the kit and load on to your PCR instrument. Post PCR all samples can be combined for pooled cleanup with the included Ampliclean Magnetic Bead Cleanup Kit and following standard quantification and QC checks your libraries are ready to sequence.

Video

Performance

Thanks to the improved reaction kinetics, intrinsic in the RC-PCR method, in combination with the well designed and balanced probe pools, the kit delivers clean data. Also, this allows for well balanced read distributions in order to make efficient use of the sequencer’s capacity. The method delivers a coverage of ~99% of the Covid Genome, and an on-target percentage of >90%. Thanks to the easy workflow, with just a few pipetting steps, this kit is the ideal solution for any virology lab, even without NGS experience. Our support team will get you up and running in no time.

Data Analysis

VirSEAK Analysis software

JSI medical systems GmbH in Germany developed a dedicated software tool for analyzing NGS data from the SARS-CoV-2 virus, with all needed settings ready installed.

Please contact JSI Medical Systems directly to learn about this software

Global GISAID data integrated

 

After the sequencing run ends, the generated fastq files can be imported into the JSI virSEAK software tool. virSEAK will automatically create a consensus sequence and alignment to the reference genome. It will show the claid where the sample belongs to and generate a .csv report of the mutations. Data will be compared to global databases (like GISAID) and you will be shown the subtype frequencies and distribution per country, as well as the evolution of the strain.

Results can be uploaded to the GISAID database with a single mouse-click in order to share the results with the global Covid community. It also provides an exportable consensus sequence for a more detailed analysis in local healthcare systems. For instance when investigating patient clusters or re-infections.

Product

EasySeq™ RC-PCR SARS CoV-2 (novel coronavirus) Whole Genome Sequencing kit (96 reactions). 

SKU: RC-COV096

 

 
Format

Generate Whole Genome Sequencing data from cDNA, derived from Covid-19 positive tested patients.

Compatibility

Compatible with any low- and mid throughput illumina® NGS sequencer of the latest generations: iSeq®, MiniSeq®, MiSeq®, NextSeq®, in combination with 2×150 bp paired-end read chemistry.

Product Information Variants Cat.
EasySeq™ RC-PCR SARS CoV-2 (novel coronavirus) Whole Genome Sequencing kit (96 reactions). 96 reactions SKU: RC-COV096
PCR 2x Hotstart HiFi Mastermix, compatible with RC-PCR assays, 2.2 mL 2 Tubes of 1.1 mL  each SKU: MMHS-25