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当前位置: 首页 > 文章 > 圆果黄麻成熟叶片总DNA提取及SRAP扩增体系的建立与优化 福建农林大学学报(自然科学版) 2011,40 (5) 461-466
Position: Home > Articles > Extraction of genomic DNA from adult jute leaves,and establishment and optimization of SRAP reaction system Journal of Fujian Agriculture and Forestry University(Natural Science Edition) 2011,40 (5) 461-466

圆果黄麻成熟叶片总DNA提取及SRAP扩增体系的建立与优化

作  者：
陈燕萍;陈美霞;徐建堂;陈晖;陶爱芬;祁建民
单  位：
福建农林大学作物遗传育种与综合利用教育部重点实验室
关键词：
圆果黄麻;DNA提取;SRAP反应体系
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.
译  名：
Extraction of genomic DNA from adult jute leaves,and establishment and optimization of SRAP reaction system
作  者：
CHEN Yan-ping1,2,CHEN Mei-xia1,XU Jian-tang1,CHEN Hui1,3,TAO Ai-fen1,QI Jian-min1(1.Key Laboratory of Ministry of Education for Genetics,Breeding and Multiple Utilization of Crops,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China;2.Agricultural Bioresource Research Institute,Fujian Academy of Agricultural Sciences,Fuzhou,Fujian 350002,China;3.Dongshan Entry-Exit Inspection and Quarantine Bureau,Zhangzhou,Fujian 363401,China)
关键词：
jute(Corchorus capsularis L.);DNA extraction;sequence-related amplified polymorphism(SRAP)
摘  要：
Genomic DNA from jute(Corchorus capsularis L.) extracted with modified CTAB method was tested for its concentration and quality by agarose gel electrophoresis and concentration detection.An optimum SRAP-PCR system was established based on optimizing the key factors of SRAP-PCR reaction system in jute.The results showed that the DNA isolated with modified CTAB method was in good quality,pure and integral.D260 nm/D280 nm value of the DNA was 1.7-1.9 using ultraviolet spectrophotometer determination.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 volume 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-1,template DNA 80-120 ng.It was showed that the optimized system in this study can be applied in the SRAP analysis for jute.

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