【创意版】RPA——PCR技术的革命.pptVIP

引物设计 RPA分析的关键在于扩增引物和探针的设计。PCR引物多半是不适用的，因为RPA引物比一般PCR引物长，通常需要达到30-38个碱基。引物过短会降低重组率，影响扩增速度和检测灵敏度。在设计RPA引物时，变性温度不再是影响扩增引物的关键因素。RPA的引物和探针设计不像传统PCR那样成熟，用户需要自己摸条件进行优化。 .......... * Speed 10 to 15 minute detection time Sensitivity Single molecule Cost Cheap access as little or no hardware is required Simplicity Stabilised reaction format, minimal sample preparation Robustness To sample contaminants and temperature fluctuations Portability Handheld instrumentation or disposable test format 为什么说RPA是一场技术革命 .......... * RPA具体应用举例 Clinical Food safety Agricultural Blood bank screening Environmental Animal health .......... * Recombinase Polymerase Amplification (RPA) of CaMV-35SPromoter and nos Terminator for Rapid Detection ofGenetically Modified Crops Chao Xu 1,?, Liang Li 1,2,?, Wujun Jin 1,2 and Yusong Wan 1,2,* 1 Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China; E-Mails: xuchao1667@163.com (C.X.); liliang@ (L.L.); jinwujun@ (W.J.) 2 Inspection and Testing Center for Environmental Risk Assessment of Genetic Modified Plant-Related Microorganisms (Beijing), Ministry of Agriculture, Beijing 100081, China .......... * 1. Introduction The International Service for the Acquisition of Agri-biotech Applications (ISAAA) estimates that millions of farmers cultivated genetically modified (GM) crops over more than 170 million hectares across 27 countries in 2013; the major GM crop species were canola, maize, cotton, and soybean [1]. Although the polymerase chain reaction (PCR) is one of the most widely used amplification methods for GMO screening detection [3], the need for delicate equipment and complicated procedures limit the use of PCR amplification in point-of-use and field settings. Rapid, specific, and highly effective methods for identifying the presence of GMOs in food and feed are important and necessary [4]. The most frequently used method for detecting GMO material is screening for the CaMV-35S promoter (P-35S) from the