野蔷薇的DNA提取和RAPD反应体系的建立

DNA Extraction and Establishment of RAPD Analytic Conditions of Rosa multiflora

以野蔷薇(Rosa multiflora)的冬末初春芽和夏初叶片为材料,研究了DNA的提取方法,并对影响随机扩增多态DNA(RAPD)的反应因素进行了优化。建立了适合野蔷薇RAPD的反应体系和反应程序,在20μL反应体系中,25ng模板DNA,2.0mmol/LMgCl2,0.2mmol/LdNTPs,1UTaqDNA聚合酶,0.1μmol/L引物。扩增程序为:94℃预变性4min,然后94℃变性45s,37℃退火1min,72℃延伸1min,37个循环,最后72℃延伸5min,4℃保存。

DNA of Rosa multiflora was extracted from the late-winter buds and the early-summer leaves. The optimized reaction system and programe of RAPD for Rosa multiflora were as follows. The reactions were performed in a volume of 20 L, which containing 25ngDNA,2. 0mmol/LMgCl2, 0. 2mmol/LdNTPs,lUTaq DNA polymerase, 0. 1 mol/L primer. The rnalcycling was programmed by 1 cycle for 5rain at 94℃ ,followed by 37 cycles for 45sec at 94℃, 1min at 37℃ ,and 1min at 72℃ and finally,by 1 cycle for 5rain at 72℃,storing at 4℃.