一种改良的转基因甘蔗基因组DNA提取方法

An Improved Method for Isolation of Transgenic Sugarcane Genomic DNA

摘要:

用改良的转基因甘蔗基因组DNA提取方法可从少量的转基因甘蔗叶片中简便快速地提取高质量的DNA,有效地去除甘蔗叶片中的多糖、多酚类和RNA等物质。经核酸蛋白测定仪及凝胶电泳分析表明,该改良方法提取的DNA具有典型的DNA分子标准紫外吸收光谱特点,其A₂₆₀/A₂₈₀为 1.7-1.9,A₂₆₀/A₂₃₀为 1.8-2.0,叶片的DNA产量为 45-60 μg(100 mg)^-1,适用于对转基因甘蔗进行PCR、酶切和Southern杂交检测分析。

Abstract:

The improved method for isolation of transgenic sugarcane genomic DNA could rapidly and simplyextract high quality DNA from a few leaves of transgenic sugarcane. The isolated DNA is essentially free of polysaccharides, polyphenols, RNA and other major contaminants. Ultraviolet absorbance spectrum analysis andgel electrophoresis results showed that DNA samples obtained by the method possessed the standard ultraviolet absorbance spectrum of pure DNA, with 1.7-1.9 A₂₆₀/A₂₈₀ and 1.8-2.0 A₂₆₀/A₂₃₀. The yield was about 45-60 μg DNA per 100 mg young leaves．The total DNA prepared by this procedure could be used for PCR, restriction enzyme digestion and southern blotting.