We have developed a novel dual enzyme chemistry called rhAmp®SNP genotyping based on RNase H2-dependent PCR (rhPCR) that provides high signal and specificity for SNP analysis. rhAmp SNP genotyping combines a u...We have developed a novel dual enzyme chemistry called rhAmp®SNP genotyping based on RNase H2-dependent PCR (rhPCR) that provides high signal and specificity for SNP analysis. rhAmp SNP genotyping combines a unique two-enzyme system with 3’ end blocked DNA-RNA hybrid primers to interrogate SNP loci. Activation of the blocked primers occurs upon hybridization to its perfectly matched target, which eliminates or greatly reduces primer dimers. A thermostable hot-start RNase H2 cleaves the primer immediately 5’ of the ribose sugar, releasing the blocking group and allowing primer extension. PCR specificity is further improved with the use of a mutant Taq DNA polymerase, resulting in improved allelic discrimination. Signal generation is obtained using a universal reporter system which requires only two reporter probes for any bi-allelic SNP. 1000 randomly selected SNPs were chosen to validate the 95% design rate of the design pipeline. A subsampling of 130 human SNP targets was tested and achieved a 98% call rate, and 99% call accuracy. rhAmp SNP genotyping assays are compatible with various qPCR instruments including QuantStudioTM 7 Flex, CFX384TM, IntelliQube®, and Biomark HDTM. In comparison to TaqMan®, rhAmp SNP genotyping assays show higher signal (Rn) and greater cluster separation, resulting in more reliable SNP genotyping performance. The rhAmp SNP genotyping solution is suited for high-throughput SNP genotyping applications in humans and plants.展开更多
DNS-based server redirecting is considered the most popular means of deploying CDNs. However, with the inaeasing use of i"emol:e DNS, DNS-based CDNs face a great challenge in performance degradation. To address t...DNS-based server redirecting is considered the most popular means of deploying CDNs. However, with the inaeasing use of i"emol:e DNS, DNS-based CDNs face a great challenge in performance degradation. To address this issue, encouraging progress has been made in both industry and research communities. In this article,state-of- art solutions for the remote DNS problem are discussed at first. Next, privacy concerns about DNS-based CDNs, including client location as well as redirection privacy, are identified and a representative solution is summarized. Finally, the solution is compared to those in prior works under different measures, and a discussion on DNS-based CDN applications is provided. A model is also established to deepen the underst;anding of CDN performance. We believe that this survey will shed light on the application of DNS-based CDNs, and it is expected to provide design guidelines to CDN service providers.展开更多
文摘We have developed a novel dual enzyme chemistry called rhAmp®SNP genotyping based on RNase H2-dependent PCR (rhPCR) that provides high signal and specificity for SNP analysis. rhAmp SNP genotyping combines a unique two-enzyme system with 3’ end blocked DNA-RNA hybrid primers to interrogate SNP loci. Activation of the blocked primers occurs upon hybridization to its perfectly matched target, which eliminates or greatly reduces primer dimers. A thermostable hot-start RNase H2 cleaves the primer immediately 5’ of the ribose sugar, releasing the blocking group and allowing primer extension. PCR specificity is further improved with the use of a mutant Taq DNA polymerase, resulting in improved allelic discrimination. Signal generation is obtained using a universal reporter system which requires only two reporter probes for any bi-allelic SNP. 1000 randomly selected SNPs were chosen to validate the 95% design rate of the design pipeline. A subsampling of 130 human SNP targets was tested and achieved a 98% call rate, and 99% call accuracy. rhAmp SNP genotyping assays are compatible with various qPCR instruments including QuantStudioTM 7 Flex, CFX384TM, IntelliQube®, and Biomark HDTM. In comparison to TaqMan®, rhAmp SNP genotyping assays show higher signal (Rn) and greater cluster separation, resulting in more reliable SNP genotyping performance. The rhAmp SNP genotyping solution is suited for high-throughput SNP genotyping applications in humans and plants.
文摘DNS-based server redirecting is considered the most popular means of deploying CDNs. However, with the inaeasing use of i"emol:e DNS, DNS-based CDNs face a great challenge in performance degradation. To address this issue, encouraging progress has been made in both industry and research communities. In this article,state-of- art solutions for the remote DNS problem are discussed at first. Next, privacy concerns about DNS-based CDNs, including client location as well as redirection privacy, are identified and a representative solution is summarized. Finally, the solution is compared to those in prior works under different measures, and a discussion on DNS-based CDN applications is provided. A model is also established to deepen the underst;anding of CDN performance. We believe that this survey will shed light on the application of DNS-based CDNs, and it is expected to provide design guidelines to CDN service providers.
