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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.

Sanger method of DNA sequencing

Sanger sequencing is a method of DNA sequencing, based on the selective incorporation of chain-terminating dideoxynucleotides by DNA polymerase during in vitro DNA replication, developed by Frederick Sanger and colleagues in 1977. Since the introduction of massive parallel sequencing (NGS), the Sanger method remains in wide use, for smaller-scale projects, validation of NGS results and for obtaining especially long contiguous DNA sequence reads.