【摘要】 对小麦黄化苗、叶片和种子基因组DNA提取,以及对影响RAPD结果的反应体系和反应条件的各项因素进行了研究。建立了快速、高效的适合于多种情况的DNA提取方法。构建了适用面宽而且稳定可靠的RAPD反应条件和反应体系。即反应条件为94℃预变性3 min,37℃退火1 min,72℃延伸2 min,循环2次,94℃变性1 min,37℃退火1 min,72℃延伸2 min,再从94℃变性1 min开始循环38次,最后72℃延伸10min;20μL反应体系为2.5 mmol/L Mg2+,0.2 mmol/L dNTP,10 pmol引物,25 ng模板,0.8 UTaq酶。更多还原

【Abstract】 Two efficient methods of extracting wheat genome DNA and an optional reaction RAPD system were established.The methods were simple and rapid in etiolations,leaves and seeds genomic DNA isolation,and the optional reaction RAPD system were set up for the assay and usage in wheat by testing and comparing some essential factors.The reaction program consisted of 2 cycles of predenaturation at 94℃ for 3 min,annealing at 37℃ for 1 min,and extension at 72℃ for 2 min,followed by 38 cycles of denaturation at 94℃ for 1 min,annealing at 37℃ for 1 min,and extension at 72℃ for 2 min.The final cycle had a 10 min extension at 72℃. The PCR mixtures of 20 μL contained 25 ng template DNA,0.2 mmol/L of each deoxynucleiotide（dNTP.dATP.dGTPandTTP）,2.5 mmol/L Mg²⁺,10 pmol primer and 0.8 U Taq polymerase.更多还原