PCR_SSCP分析条件的优化.pdfVIP

( ) 31 1 Journal of Fujian Agriculture and Forestry University( Natural Science Edition) 2002 3 - 分析条件的优化 PCR SSCP , , ( 福建农林大学植物病毒研究所, 福建 福州350002) : ( PCR-SSCP) , . PCR-SSCP , . : PCR-SSCP; : S435 : A : 1006-7817(2002) 01-0022-04 - Optimization of the conditions affecting PCR SSCP analysis WEI T ai-yun, IN Han-xin, XIE ian-hui (Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China) : - - ( - ) Abstract Polymerase chain reaction single strand conformation polymorphism PCR SSCP technique is a method feasible for detect ing minor sequence changes in PCR amplified DNA, with its characterization of rapidness, simplicity, sensitivity - . , , and applicability to large scale screening However many factors can influence the result s of SSCP analysis and it s opti- mization is highly empirical. In this paper, w e compared and analyzed the experiment al results under various controlled - , , conditions such as the concentration of cross linker the concentration of PAG the concentration of the electrophoresis buffer, the presence of glycerol in gels and temperature of the gel during electrophoresis. Results show ed that it was com- mon t o run the samples under at least two different conditions because each fragment needed it s ow n opt imal running condi- tions. : - ; Key words PCR SSCP conditions optimization (polymerase chain reaction-single strand conformation polymor- , - ) . phism PCR SSCP PCR