摘要
[目的]以嗜酸乳杆菌(Lactobacillus acidophilus)为试验菌株,寻找一种高质量、高稳定性的细菌基因组DNA的提取方法。[方法]采用球磨-十六烷基三甲基溴化铵(Cetyltrimethyl ammonium bromide,CTAB)法提取细菌基因组DNA,通过调整研磨珠直径、菌体量、破碎时间三因素对DNA提取效果进行正交优化。[结果]确定最佳优化条件为破碎时间3.5 min、菌体量9×10~8CFU/m L、研磨珠直径0.1 mm。[结论]经条件优化后的球磨-CTAB法短时间内可获得高质量的细菌DNA产品,处理量大、方便快捷。
[Objective] L. acidophilus was used as the experimental strain to find a high quality and high stability bacterial genomic DNA extraction method. [Methods] The DNA was extracted by the beadbeater-CTAB method and the DNA extraction efficiency was optimized orthogonally by adjusting the diameter of grinding beads,the amount of bacteria and the beating time.[Results]The best optimum conditions for the final determination were 3. 5 min in break time,9 × 10^8 CFU/m L in bacterial cells and 0. 1 mm in diameter. [Conclusion] High-quality DNA products can be obtained at short time by the optimized Beadbeater-CTAB method,and the processing capacity is large and convenient.
作者
臧凯丽
王泳
赵林森
闫亚丽
崔文静
马新颖
赵培
陈庆森
Kaili Zang;Yong Wang;Linsen Zhao;Yali Yan;Wenjing Cui;Xinying Ma;Pei Zhao;Qingsen Chen(Tianjin Key Laboratory of Food Biotechnology, College of Biotechnology and Food Science, Tianjin University of Commerce, Tianjin 300134, China;Hebei Inatural Biological Technical Company, Shijiazhuang 050800, China)
出处
《生物技术》
CAS
2018年第3期236-241,共6页
Biotechnology
基金
国家自然科学基金面上项目("酪蛋白糖巨肽(CGMP)维系肠粘膜免疫自稳机制的研究"
No.31071522)
关键词
球磨-CTAB法
提取
嗜酸乳杆菌
基因组DNA
正交优化
the beadbeater - CTAB method
extraction
Lactobacillus acidophilus
genome DNA
orthogonal optimization
