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以石榴(Punica granatum L.)成熟叶片为试材,设置6种保存方法,采用改良的CTAB法提取石榴基因组DNA,研究了不同保存方法对石榴叶片基因组DNA提取效果的影响。结果表明:采用改良的CTAB法,新鲜叶片、-70℃保存6个月、4℃保存5d的样品均能得到浓度较高的DNA;-20℃保存7d和14d的样品均能得到完整DNA,但浓度略低;硅胶干燥常温保存5个月的样品用同样的方法未能提取到DNA,在对提取方法进行优化后,可提取纯度较高的基因组DNA,但其浓度仅约为鲜叶的50%。上述方法保存的样品提取的基因组DNA均能得到清晰、稳定的SRAP-PCR扩增图谱。
The mature leaves of Punica granatum L. were used as materials to set up six kinds of preservation methods. The genomic DNA of pomegranate was extracted by modified CTAB method. The effects of different preservation methods on the extraction of genomic DNA from pomegranate leaves were studied. The results showed that the higher concentration of DNA could be obtained by modified CTAB method with fresh leaves at -70 ℃ for 6 months and at 4 ℃ for 5 days. The intact DNA could be obtained from the samples stored at -20 ℃ for 7d and 14d, But the concentration was slightly lower. The sample that was stored at room temperature for 5 months on silica gel failed to extract DNA by the same method. After the extraction method was optimized, the genomic DNA with higher purity could be extracted, but its concentration was only about 50%. The genomic DNA extracted from the samples preserved in the above method can obtain a clear and stable SRAP-PCR amplification map.