摘要
采用改良的CTAB法从圆果黄麻成熟叶片中提取基因组DNA,对DNA进行电泳检测、含量测定和SRAP分析,并对黄麻SRAP-PCR反应体系中主要影响因子进行了优化,建立了最佳反应体系.结果表明,改良的CTAB法能够提取高质量的DNA,DNA纯度和完整性都较好,经紫外分光光度计测定,D260 nm/D280 nm均在1.7-1.9之间,无降解现象,蛋白质、多糖类等去除彻底.所建立的SRAP最佳反应体系(25μL)中5种成分的适宜浓度及用量分别为:Taq DNA聚合酶1.75 U,Mg2+3.0 mmol.L-1,dNTPs 0.16 mmol.L-1,引物0.51μmol.L-1,模板DNA 80-120 ng.
C, enomie DNA from jute （ Corchorua capsularis L. ） extracted with modified CTAB method was tested for its concentration and quality by agarose gel electrophoresls and concentration detection. An optimum SRAP-PCR system was established based on optimizing the key factors of SRAP - PCR reaction system in jut~. The results showed that the DNA isolated with modified CTAB meth- ed was in good quality, pure and integral. D260nm/D280nm value of the DNA was 1.7 - 1.9 using ultraviolet spectropbetometer deter- mination. There was no degradation appeared and the proteins, phenols, etc. were eliminated completely. The optimum SRAP-PCR reaction system could amplify high levels of polymorphism, had good repeatability and clear band pattern. SRAP system （ total vol- ume of 25 μL） was established as follows： Taq DNA polymerase 1.75 U, Mg2＋3.0 mmol· L-1 , dNTPs 0.16 mmol· L-1, forward primer 0.51 μmol· L^-1 , reverse primer 0.51 μmol · L-l , template DNA 80 - 120 ng. It was showed that the optimized system in this study can be applied in the SRAP analysis for jute.
出处
《福建农林大学学报（自然科学版）》
CSCD
北大核心
2011年第5期461-466,共6页
Journal of Fujian Agriculture and Forestry University:Natural Science Edition
基金
国家自然科学基金(30571188)资助项目
国家麻类产业技术体系建设项目(nycytx-19-E05)
关键词
圆果黄麻
DNA提取
SRAP反应体系
jute （ Corchorus capsularis L. ）
DNA extraction
sequence-related amplified polymorphism （SRAP）
