摘要
以延龄草基因组DNA为模板,对ISSR反应体系中的一些重要参数进行了探索和优化实验,确立了适合延龄草的ISSR反应体系,即20μL反应体积中,模板DNA为20 ng、引物浓度为0.6μM、dNTP浓度为0.6 mM、Taq DNA聚合酶1 U.反应循环过程为:94℃变性30 s、52℃退火45 s、72℃延伸2 min、45个循环,72℃延伸7 min.该体系重复性和稳定性都较好,可有效地应用于延龄草遗传多样性的研究.
Based on the genomic DNA extracted from Trillium tschonoskii Maxim, some essential factors that might affect the result of ISSR assay were tested and compared. An optimal reaction system suitable for the assay and usage of ISSR was established. Amplification reactions contained 20 ng genomic DNA, 0.6 μM primers, 0.6 mM of each dNTP and 1.0 unit Taq DNA polymerase in a volume of 20μL. The reaction program was devised for 45 cycle, each with 30 s at 94 ℃, 45 s at 52 ℃, 2 min at 72 ℃ and 7 min at 72 ℃ in final extension. This system with good reproducibility and high reliability could be effectively used for ISSR analysis in the program of Trillium tschonoskii Maxim genetic diversity.
出处
《四川师范大学学报（自然科学版）》
CAS
CSCD
北大核心
2007年第4期523-525,共3页
Journal of Sichuan Normal University（Natural Science）
基金
国家自然科学基金(30300023)资助项目
关键词
延龄草
ISSR反应体系
遗传多样性
Trillium tschonoskii Maxim
ISSR reaction system
Genetic diversity
