实验目的
对真菌细胞进行破壁,提取其DNA。
实验原理
随着分子生物学技术的进展,PCR相关方法越来越多地被应用到真菌学研究中来,而真菌DNA的提取是包括分子诊断在内的所有实验的基础,简便快捷地提取真菌DNA,且获得的DNA完整、纯度高,对真菌分子生物学的研究有及其重要的意义。
真菌细胞壁以几丁质、葡萄糖为主构成细胞壁骨架,而基质则由多糖、甘露聚糖、糖蛋白、脂质等填充。丝状真菌坚韧厚壁的构成,需要较为繁琐的破壁手段,故真菌DNA的提取较细菌或其他组织细胞难度更大。为保证真菌PCR技术及其相关分子生物学技术的顺利开展,简便快捷地提取真菌DNA,且获得的DNA完整、纯度高,是必须解决的前提条件。
实验材料和器具
样品:真菌类
仪器:高通量组织研磨仪(上海净信,Tissuelyser-48)
耗材:离心管,研磨珠(上海净信)
试剂:TE Buffer,DA Buffer,E Binding Buffer,Elution Buffer
实验步骤
需了解具体步骤,请致电厂家
实验结果
图1 提取DNA的检测 M:100bp 标准参照物
图2 提取DNA的PCR检测结果 M:100bp 标准参照物
Previous:高通量组织研磨仪对多酚含量高的植物(木本植物)的细胞破壁
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