丝瓜issr-pcr反应体系的优化.pdf

福建农业学报26( 1) : 70~ 75 , 2011 : 1008- 0384 (2011) 0 1- 70- 0 6 Fuj ian Journal of Agricultural Sciences ISSR-P CR 1 2 2 3 2 陈朝文, 林 辉 , 薛珠政 , 林义章 , 温庆放 ( 1. , 362200 , 2. , 350018; 3. , 35000 2) : , ISSR- PCR , : 20 L , 20 L 10 bu er , 15 U TaqDN A , 025 mmo l L- 1 dN T Ps, 035 mo l L - 1 , 30 ng DN A , ISSR- PCR CW32505 541 : ; ISSR; : S 6424 : A Optimization of ISSR-PCR reaction system for Luffa 1 2 2 3 2 CH EN Chao- w en , L IN Hui , XU E Zhu- zheng , L IN Y-i zhang , WEN Q ing- ang (1. Quanzhou institute of Agricultural Sciences, Quanz hou, Fuj ian 362200, China; 2. Crop Research I nstitute, Fuj ianAcade y of A gricultural Sciences Fuzhou, Fuj ian 3500 18, China; 3. College of H orticulture, Fuj ianA griculture and Forestry University , Fuz hou, Fuj ian 350002, China) Abstract: T o di erentiate v ariou s lu a po pulat ions and or g enetic relationship analy sis among them , their tender leaves w ere used in this experiment to establish the reactio n system . Factor s a ecting the ISSR- PCR ampli ication w er e selected and optim ized thro ug h the combinatio n o a sing le acto r and or thogo nal design ex periment . A stable and repro ducible r eaction system suitable or ISSR- P CR analysis o r lu a containing 20 L 10 bu er, 15 U TaqDN A po lymer ase, 025 mmol L- 1 dN T P s, 035 mo l L- 1 primer and 30 ng templat e DN A per 20 L