圆果黄麻成熟叶片总DNA提取及SRAP扩增体系的建立与优化

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,D260nm/D280nm均在1.7-1...

Extraction of genomic DNA from adult jute leaves, and establishment and optimization of SRAP reaction system

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.