HAO Dou-dou,ZHU Yong,LEI Ming,et al.Comparison of Different Extraction Methods of Genomic DNA from Avena sativa L[J].HEILONGJIANG AGRICULTURAL SCIENCES,2015,(10):20-23.[doi:10.11942/j.issn1002-2767.2015.10.0020]
燕麦种子基因组DNA不同提取方法的比较
- Title:
- Comparison of Different Extraction Methods of Genomic DNA from Avena sativa L
- 文章编号:
- 1002-2767(2015)10-0020-04
- Keywords:
- Avena sativa L seed; extraction method; PCR amplification
- 分类号:
- S543+.7
- 文献标志码:
- A
- 摘要:
- 为缩短DNA提取时间,省去种子萌发到幼苗培养等一系列过程,以燕麦种子为材料,用5种方法提取其基因组DNA,通过紫外分光光度法检测DNA的浓度和纯度,琼脂糖凝胶电泳检测DNA的完整性,以ITS2为引物将燕麦种子基因组DNA进行PCR扩增。结果表明:5种方法中除了传统CTAB法,其余方法都可以提取到燕麦种子基因组DNA,但不同方法提取到的基因组DNA的纯度、浓度存在差异,试剂盒法提取的DNA质量最好、纯度最高,但提取的DNA量少且成本高,改良CTAB法和高盐低pH法提取的DNA的纯度相近,都有少量的蛋白质和糖类的污染,改良SDS法提取的DNA纯度最低。
- Abstract:
- In order to overleap seeding culture and decrease the experimental time,taking Avena sativa L seeds as material,the genomic DNA of Avena sativa L were extracted by five kinds of methods.The concentraction and purity of DNA were detected by ultraviolet spectrophotometry.And integrity was detected by agarose gel electrophoresis.Based on ITS2 for primers,the DNA was detected through PCR amplification.The results showed that except for traditional CTAB,the other four methods could extract the genomic DNA from Avena sativa L seeds,there were obvious difference in extracted purity of genomic DNA with different methods,DNA was extracted with the kit method which had the highest quality and purity,but it was expensive and had less DNA extracted.The difference of DNA purity between improved CTAB method and high salt and low pH method was not very obvious,there was a small number of protein and carbohydrate in DNA.The purity of DNA was the lowest with improved SDS method.
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备注/Memo
收稿日期:2015-07-19
基金项目:湖南师范大学蛋白质与发育生物学教育部重点实验室(2014年)开放课题资助项目(2015DF03);西藏自治区科技厅自然科学基金资助项目(2015ZR135)
第一作者简介:郝豆豆(1992-),女,山西省吕梁市人,在读硕士,从事植物学研究。E-mail:31724478@qq.com。
通讯作者:拉多(1968-),男,藏族,博士,副教授,硕士生导师,从事藏药材基因与功能研究。E-mail;lhaduo@hotmail.com。