摘要
以南瓜矮生和蔓生的自交系为材料,通过对影响南瓜AFLP反应体系中的关键因素(如DNA的提取方法、酶切和连接反应时间、预扩增产物稀释倍数和银染检测方法等)进行研究,结果表明,采用加入PVP和β-巯基乙醇后的改良CTAB提取法获得的南瓜嫩叶基因组DNA质量较好;5.5 h可以满足酶切和连接反应;酶切连接产物稀释5倍,预扩增产物稀释30倍,可以满足试验要求。应用该体系,对96对引物进行筛选,在南瓜矮生和蔓生自交系间获得的引物扩增出了重复性好、清晰的南瓜矮生和蔓生的自交系多态性条带。为进一步进行南瓜矮生基因定位奠定了良好的基础。
Based on dwarf and vine inbred lines,key factors of AFLP analysis system（such as genomic DNA extraction,time requirement of digestion and ligation reactions,dilution of pre-amplification and silver staining method et al） were studied.The result showed： the best quality genomic DNA were adding PVP and β-mercaptoethanol modified CTAB method;5.5 h can be meet digestion and ligation reactions;Diluted 5 times for products of digestion and ligation reactions and diluted 30 times for products of pre-amplification can be meet experimental conditions.96 primers screened,reproduce,clear polymorphic bands were obtained between dwarf and vine inbred lines.It laid good foundation to tropic pumpkin dwarf vine mapping.
出处
《华北农学报》
CSCD
北大核心
2011年第2期124-127,共4页
Acta Agriculturae Boreali-Sinica
基金
山西省回国留学人员科研项目(20040872010112)
关键词
南瓜
矮生
AFLP体系
Tropic pumpkin
Dwarf
AFLP analysis system
