湖北农业科学
湖北農業科學
호북농업과학
2010年
3期
522-525
,共4页
兰小平%李三相%雒林通%李一婧%王廷璞%李燕%徐丽%鄢殉%崔泰保
蘭小平%李三相%雒林通%李一婧%王廷璞%李燕%徐麗%鄢殉%崔泰保
란소평%리삼상%락림통%리일청%왕정박%리연%서려%언순%최태보
藏獒%DNA提取%随机扩增多态性DNA%体系优化
藏獒%DNA提取%隨機擴增多態性DNA%體繫優化
장오%DNA제취%수궤확증다태성DNA%체계우화
Tibetan mastiff%DNA extraction%random amplified polymorphic DNA (RAPD)%system optimization
采用改进的盐析法提取了藏獒血液基因组DNA,并对RAPD扩增体系进行了优化.结果表明,改进的盐析法所提取的藏獒基因组DNA完整、无降解,完全可以满足RAPD分析的需要;20 μL的最佳扩增体系组成为2.50mmoL/L Mg~(2+)、0.60mmoL/L dNTP、50 ng模板DNA、1.25U Taq酶和0.3μmol/L随机引物,最佳退火温度为36.5℃.
採用改進的鹽析法提取瞭藏獒血液基因組DNA,併對RAPD擴增體繫進行瞭優化.結果錶明,改進的鹽析法所提取的藏獒基因組DNA完整、無降解,完全可以滿足RAPD分析的需要;20 μL的最佳擴增體繫組成為2.50mmoL/L Mg~(2+)、0.60mmoL/L dNTP、50 ng模闆DNA、1.25U Taq酶和0.3μmol/L隨機引物,最佳退火溫度為36.5℃.
채용개진적염석법제취료장오혈액기인조DNA,병대RAPD확증체계진행료우화.결과표명,개진적염석법소제취적장오기인조DNA완정、무강해,완전가이만족RAPD분석적수요;20 μL적최가확증체계조성위2.50mmoL/L Mg~(2+)、0.60mmoL/L dNTP、50 ng모판DNA、1.25U Taq매화0.3μmol/L수궤인물,최가퇴화온도위36.5℃.
The method of improved salting out was used for whole blood genomic DNA extraction of Tibetan mastiff, and theRAPD reaction system of Tibetan mastiff was optimized. The results showed that the integrity and non-degradable genomicDNA of Tibetan mastiff could be gained by the method of improved salting out, and a total volume of 20 μL optimal RAPDreaction system of Tibetan mastiff consisted of 2.50 mmol/L Mg~(2+), 0.60 mmol/L dNTP, 50 ng genomic DNA, 1.25 U TaqDNA polymerase, and 0.3 μol/L arbitrary primers. The best renaturation temperature of RAPD analysis of Tibetan mastiffwas 36.5℃.
