摘要
采用改进的SDS DNA提取方法从梅花的嫩叶和老叶中提取获得总DNA .所得DNA样品的OD2 6 0 OD2 80 值在1 7～ 1 9之间 ,琼脂糖凝胶上主带清晰、较少降解、样品纯度高、蛋白去除干净且得率高 .该体系对原提取程序作了多处简化 ,使操作简便、快捷、高效 ,适合于梅花DNA提取 ,所提取的DNA样品不含PCR反应抑制剂及其他酶反应抑制剂 ,可被限制性内切酶MseⅠ和EcoRⅠ完全酶切 ,适合AFLP PCR反应应用 ,得到清晰的指纹式样 .
The improved method of SDS DNA isolation is used to extract total DNA from tender or old leaves of \%Prunus mume\% in this study. High quality total DNA from plant was isolated in less than 3 hours. The values of OD 260 / OD 280 of each sample were between 1 7 and 1 9 Strips in agarose electrophoresis were clear and less decomposed. DNA samples with high purification and high yield rate could be gained by this method. It is much more simplified than former methods at many steps. The procedure of the method is easy, fast and effective, and is good to extract genomic DNA of \%Prunus mume\%. These genomic DNA samples are intact without contaminating nucleases or inhibitor. The DNA isolated with this method could be completely digested by restriction enzyme of Mse Ⅰ and EcoR Ⅰ, and good enough for use in following applications including PCR and analysis of AFLP. The systems suitable for silver stained AFLP analysis were established and capable of producing sharp fingerprint patterns.
出处
《北京林业大学学报》
CAS
CSCD
北大核心
2003年第3期17-21,共5页
Journal of Beijing Forestry University
基金
湖北省自然科学基金资助项目 ( 2 0 0 1ABB118)
关键词
梅花
DNA提取
AFLP
DNA模板
银染
Prunus mume, DNA isolation, AFLP, DNA template, silver staining
