摘要
以改良CTAB法从猕猴桃叶片中制备模板DNA ,优化了PCR热循环参数 ,建立了RAPD PCR扩增的最佳反应体系。实验结果表明 ,CTAB提取液中EDTA组分的浓度对模板提取影响很大 ,其最适浓度为 80mmol/L ;用异丙醇沉淀后不经乙醇洗涤纯化的DNA不会影响扩增效果。PCR热循环参数为 :94℃预变性 5min ;94℃变性 1min ,37℃退火 1min ,72℃延伸 2min ,循环 4 0次 ;最后在 72℃延伸 6min。
The revisory method CTAB was used to extract DNA from Actinidia plant-leaves. The recycling parameters were optimized and then optimal reaction system suitable for RAPD-PCR amplification was established. The results show that the element EDTA in the solution for extraction effected greatly on the abstraction of genomic DNA, which is suitable by 80mmol/L. The amplification wasn't be effected by the template which wasn't purified by the ethanol. The RAPD-PCR reaction program is composed of an initial heat denaturation for 5 min at 94℃, the amplification for 40 cycles, consisting of 1min at 94℃, 1 min at 37℃, 2 min at 72℃, and the final extension which is 6 min at 72℃.
出处
《生物技术通报》
CAS
CSCD
2003年第3期40-43,共4页
Biotechnology Bulletin
基金
国家自然科学基金资助项目 ( 3 9960 0 0 7)
江西省自然科学基金资助项目 ( 983 0 2 7)
