机构地区: 华南农业大学园艺学院
出 处: 《中国农学通报》 2010年第6期22-25,共4页
摘 要: 利用改良的CTAB方法从芥蓝叶片中提取高质量的DNA。在参考一般RAPD分析反应程序的基础上,经过优化试验,确定适合芥蓝PCR扩增体系(总体积25μL)为:25mmol/LMgCl22.0μL、10×PCR Buffer2.5μL、2.5mmol/LdNTPs2.0μL、5U/μLTaq酶0.25μL、5μmol/L引物1.2μL、模板DNA25ng、灭菌双蒸水12.25 μL。PCR扩增程序为:94 ℃预变性5 min,94℃变性1 min,36 ℃退火1 min,72 ℃延伸2 min,40个循环,72 ℃延伸10 min。 High quality DNA was obtained from Chinese Kale (Brassiea alboglabra Bailey) leaf by the improved method of CTAB. Based on the common RAPD reaction program and the adjusting experiments, the optimal PCR system (25 μ L total volumes)contains: 5 mmol/LMgC12 2.0μ L, 10× PCR Buffer 2.5 μ L, 2.5 mmol/L dNTPs2.0 μ L, 5U/μ L Taq DNA polymerase 0.25μ L, 5 μ mol/L primerl.2 μ L, genomic DNA 25ng and ddH2O 12.25μ L. The optimal amplification program as follows: 94℃ for 5 min; 40 cycles at 94℃ for 1 rain, 36℃ for 1 rain and 72℃ for 2 rain; 72℃ for 10 min at last. :