不同保存方法和提取方法对扁桃基因组DNA质量的影响

以扁桃（Amygdalus conmmuis L.）幼叶为试材,通过7种保存方法并采用改良2×CTAB法、3×CTAB法和SDS法对其DNA进行提取,用琼脂糖凝胶电泳检测其DNA完整性,分光光度计测其产量和纯度,以探讨不同保存方法和不同提取方法对扁桃DNA提取质量的影响。结果表明,扁桃新鲜幼叶、压干幼叶后在-75℃保存、45℃烘干幼叶后在-75℃保存以及硅胶干燥幼叶后在-75℃保存三个月均可以用三种提取方法可获得完整的DNA,纯度和产量均高,其保存和提取方法简便易行且获得高质量DNA。扁桃鲜叶直接放在-20℃保存比-75℃保存更易获得DNA,而压干后的扁桃叶片-20℃保存三个月并采用SDS法提取,获得高产量、高纯度的DNA,不影响提取效果。SDS法和改良3×CTAB法均可获得扁桃DNA,但SDS法优于3×CTAB法,其操作简单、耗时少;改良2×CTAB法提取扁桃成功率较低,不稳定,产量比其它两种方法低。

Effects of Different Sample Preserving Methods and Extracting Methods of Genomic DNA of Almond

Experiment was conducted with young leaves of Amygdalus conmmuis L.and 3 methods,including modified 2×CTAB,3×CTAB and SDS were used for total DNA of almond.The samples were stored 7 methods for 3 months and the integrity,yield and purity of total genomic DNA were examined by UV-spectrophotomoter and agarose gel electrophoresis in order to make clear how the preserving and collection methods affecting the DNA quality.The results showed that total DNA extracted from fresh young almond leaf,pressured-dried leaf at-75℃,dried leaf at 45℃ and silica gel-dried leaf at-75℃ for three months with three extracting methods and the purity and yield of almond DNA are high,while the four preservations are convenient and ideal.The high-quality almond DNA was extracted more easily from leaf preserved at-20℃ than at-75℃ after three months,and the pressured-dried leaf preserved at-20℃was suited to obtain their highly-quality DNA using SDS method.Overall modified 3×CTAB and SDS were all good extracting almond DNA,but SDS method is the more optimum than the 3 ×CTAB under a variety of preservation conditions,work better at extracting high-quality genomic DNA.SDS method is easy to be operated,costs less time and superior to the 3 ×CTAB method.High-quality DNA used 2 ×CTAB method can not be always obtained steadily and the yield was lower than the other methods form almond leaf.