【摘要】 为研究湖南宽皮柑橘的血缘与起源关系,以湖南宽皮柑橘为试验材料,探讨其SRAP分子标记的反应体系.结果表明:设计并筛选了110对引物,其中28对引物扩增带型丰富,以引物组合Me5-Em11、Me9-Em10多态性最高,研究确定在25μL的反应体系中,Mg2+浓度在1.5 mmol.L-1,dNTP 0.3 mmol.L-1,上下引物各为0.2μmol.L-1,Taq酶浓度为0.3 U效果好,最佳PCR程序为94℃预变性5 min,然后94℃,1 min;50℃,1 min;72℃,1 min,35个循环,最后72℃延伸10 min,利用优化后的反应体系成功构建了湖南宽皮柑橘的指纹图谱.更多还原

【Abstract】 With a purpose to clarify the relationships among the native citrus resources in Hunan,this paper developed an SRAP marker reaction system for mandarins.The results show that clean and stable bands are detected for twenty-eight pairs of SRAP primer which have been selected from 110 pairs;that the best polymorphic bands are produced for pairs of Me5-Em11 and Me9-Em10 primers;that each 25 μL PCR reaction mixture consists of genomic DNA,1.5 mmol·L-1 of MgCl2,0.3 mmol·L-1 of dNTP,0.2 μmol·L-1 of primers and 0.3 U unit of Taq polymerase;and that this mixture is initially denatured at 94 ℃ for 5 min,then rotated 35 times at 94 ℃ for 1 min,50℃ for 1 min,and 72 ℃ for 1 min,and finally extended at 72 ℃ for 10 min.With this reaction system being optimized, a genetic linking map of mandarins(Citrus reticulata Blanco) in Hunan has been established.更多还原