Designing primers for PCR-based diagnostics was achieved by executing sight searches on DNA sequences. Visual searching for specific DNA targets is time consuming, subjective and requires optimisation among numerous c...Designing primers for PCR-based diagnostics was achieved by executing sight searches on DNA sequences. Visual searching for specific DNA targets is time consuming, subjective and requires optimisation among numerous candidate primer sets. Several primer design software have been linked to useful bioinformatic packages to speed the development of PCR assays. Despite the software options available, primer design has remained a challenging aspect of incursion responses, biosecurity emergencies and microbial forensic applications. Two surveys were conducted among 45 plant virologists and 21 other plant pathologists during the 7th Australasian Plant Virology Workshop and the 16th Biennial Australasian Plant Pathology Conference in 2006 and 2007, respectively. Results show that most primer design learning occurs scientist to scientist rather than during academic teaching. This tendency matches with 16% of scientists users of PCR, who do not engage in primer design and 25% designing primers only by visual means, combining a pool of 41% who if trained, would likely enhance their performance in primer design. Only 13 out of 58 scientists ranked themselves as experts. Implementing primer design in study programs and regional training will benefit plant pathology and entomology, and the responsiveness and performance of biosecurity and microbial forensics in the South Pacific.展开更多
针对超高速射武器弹药对高频高安全及高一致性点火的发展需求,基于半导体桥芯片点火技术,设计了一种新型钝感高瞬发底火。通过对比研究4种半导体桥芯片及其2种封装结构,选用斯蒂芬酸铅和亚铁氢化铅/高氯酸钾作为两级复合装药,优化形成...针对超高速射武器弹药对高频高安全及高一致性点火的发展需求,基于半导体桥芯片点火技术,设计了一种新型钝感高瞬发底火。通过对比研究4种半导体桥芯片及其2种封装结构,选用斯蒂芬酸铅和亚铁氢化铅/高氯酸钾作为两级复合装药,优化形成了模块化结构的半导体桥底火样机。该底火满足了与弹药接口的匹配要求,并达到1 A 1 W 5 min不发火的安全性要求。发火性能试验表明,所研制的半导体桥底火作用时间不大于200μs,散布不大于30μs,与传统桥丝电底火相比瞬发度和作用时间一致性显著提高,综合性能更好。展开更多
[Objective] This study aimed to reconstruct the HP intein by overlap-exten- sion PCR. [Method] Several primers were designed according to the principle of overlap-extension PCR that the repeat sequences were overlappe...[Objective] This study aimed to reconstruct the HP intein by overlap-exten- sion PCR. [Method] Several primers were designed according to the principle of overlap-extension PCR that the repeat sequences were overlapped and served as the template to the other DNA strand in amplification. Then, several rounds of over- lap PCR reaction were conducted to mutate the four cysteines in the HP intein into serines, and introduce a linker sequence containing a tag for product detection and purification. [Result] DNA sequencing analysis proved that the four cysteines in the HP intein were successfully mutated into serines; the PCR precursor fragments were also rejoined into an intact HP gene fragment, without introducing any other unex- pected mutation; the DNA linker of 46 bp was successfully inserted into the de- signed HP gene sequences, without any mutation and base pair mismatch. [Conclu- sion] The HP intein was rapidly reconstructed by overlap-extension PCR in this study, which not only expands the application of overlap PCR in protein engineering, but also provides a simple, efficient and highly accurate tool for the reconstruction and modification of intein.展开更多
文摘Designing primers for PCR-based diagnostics was achieved by executing sight searches on DNA sequences. Visual searching for specific DNA targets is time consuming, subjective and requires optimisation among numerous candidate primer sets. Several primer design software have been linked to useful bioinformatic packages to speed the development of PCR assays. Despite the software options available, primer design has remained a challenging aspect of incursion responses, biosecurity emergencies and microbial forensic applications. Two surveys were conducted among 45 plant virologists and 21 other plant pathologists during the 7th Australasian Plant Virology Workshop and the 16th Biennial Australasian Plant Pathology Conference in 2006 and 2007, respectively. Results show that most primer design learning occurs scientist to scientist rather than during academic teaching. This tendency matches with 16% of scientists users of PCR, who do not engage in primer design and 25% designing primers only by visual means, combining a pool of 41% who if trained, would likely enhance their performance in primer design. Only 13 out of 58 scientists ranked themselves as experts. Implementing primer design in study programs and regional training will benefit plant pathology and entomology, and the responsiveness and performance of biosecurity and microbial forensics in the South Pacific.
文摘针对超高速射武器弹药对高频高安全及高一致性点火的发展需求,基于半导体桥芯片点火技术,设计了一种新型钝感高瞬发底火。通过对比研究4种半导体桥芯片及其2种封装结构,选用斯蒂芬酸铅和亚铁氢化铅/高氯酸钾作为两级复合装药,优化形成了模块化结构的半导体桥底火样机。该底火满足了与弹药接口的匹配要求,并达到1 A 1 W 5 min不发火的安全性要求。发火性能试验表明,所研制的半导体桥底火作用时间不大于200μs,散布不大于30μs,与传统桥丝电底火相比瞬发度和作用时间一致性显著提高,综合性能更好。
基金Supported by the National High Technology Research and Development Program of China(863Program)(2006AA03Z451)Shanghai Basic Research Key Program(10JC1400300)+1 种基金National Natural Science Foundation of China(31070698)the Fundamental Research Funds for the Central Universities(12D10520)~~
文摘[Objective] This study aimed to reconstruct the HP intein by overlap-exten- sion PCR. [Method] Several primers were designed according to the principle of overlap-extension PCR that the repeat sequences were overlapped and served as the template to the other DNA strand in amplification. Then, several rounds of over- lap PCR reaction were conducted to mutate the four cysteines in the HP intein into serines, and introduce a linker sequence containing a tag for product detection and purification. [Result] DNA sequencing analysis proved that the four cysteines in the HP intein were successfully mutated into serines; the PCR precursor fragments were also rejoined into an intact HP gene fragment, without introducing any other unex- pected mutation; the DNA linker of 46 bp was successfully inserted into the de- signed HP gene sequences, without any mutation and base pair mismatch. [Conclu- sion] The HP intein was rapidly reconstructed by overlap-extension PCR in this study, which not only expands the application of overlap PCR in protein engineering, but also provides a simple, efficient and highly accurate tool for the reconstruction and modification of intein.
