| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | The DSN gene was synthesized by Genscript (NJ, USA) with an N-terminal 6X His tag followed by a TRX tag to facilitate disulfide bond formation. Three mutant DSN enzymes were created [1) D361N;2) D362N; and 3) D361N, D362N]via site directed mutagenesis and confirmed by Sanger sequencing. Mutant DSN enzymes were expressed in SHuffle T7 Express Competent E. coli cells (New England Biolabs, MA, USA), to enhance disulfide bond formation and proper protein folding. Shuffle cells were grown in Overnight Express at 28°C for 18 hours and harvested at 12,000g for 20 minutes at 4°C. | Get A Quote |
RNA viruses are a potent human adversary, evidenced by several global pandemics including the Ebolavirus in West Africa, the emerging Zika virus, and outbreaks of new Influenza strains and Norwalk virus in the food supply and cruise ships. Despite the virulence of these pathogens, there remains a significant limitation for detecting these viruses in a fast, accurate and cost effective manner. To meet this need we present a modified form of the duplex specific nuclease enzyme from the Paralithodes camtschaticus crab capable of generating an RNA-based signal amplification in a fraction of the time required for standard RT-qPCR. The applicability of this enzyme is demonstrated in an assay for Norwalk virus detecti... More
