RAPD条件优化及天麻基因组DNA多态性分析

Optimizing of RAPD Amplifying Condition and Analysis of Genetic DNA Polymorphisms of Gastrodia elata Blume

建立了RAPD扩增条件快速优化程序与方法,并应用于天麻基因组DNA扩增条件的优化及多态性的测定;获得了天麻基因组DNA的RAPD扩增优化条件和DNA指纹图谱;分析了模板DNA、引物、dNTP、TaqDNA聚合酶等的浓度和退火温度对RAPD扩增的影响.结果表明:天麻基因组DNA用引物S1扩增的片段具有更明显的多态性,这种指纹图谱更适合于天麻遗传分化研究;而用引物S12扩增的DNA指纹图谱具有更大的相似性,这种指纹图谱更适合于天麻真伪鉴别.该方法使RAPD扩增条件优化过程实现了程序化和数量化,是获得RAPD优化条件的简便快速、经济实用方法.应用该方法进行RAPD扩增,可获得图谱清晰、稳定可靠的实验结果.

The method protocol of optimizing rapidly RAPD amplifying condition i s set up and it is applied in optimizing amplifying conditions of the genomic DN A of Gastrodia elata Blume and determining of their DNA polymorphisms.The optimi zed conditions of the RAPD amplification of the genomic DNA of Gastrodia elata B lume and their DNA fingerprints are obtained.The influences were analyzed from c oncentration of template DNA,primer,Taq DNA polymerases,dNTPs and so on,annealin g temperatures on RAPD amplification.The results showed the fragments of amplify ing the genetic DNA of Gastrodia elata Blume with the primer S1 have more polymo rphisms.The type of DNA fingerprints still more suit the study of genetic diver gence in Gastrodia elata Blume.But the DNA fingerprints amplified with the prim er S12 have a greater similarity;the type of fingerprints still more suits the d iscrimination of true and false Gastrodia elata Blume.The method makes the optimizing process of RAPD amplification conditions a procedure and optimizing conditions expressed with quantity.It is a handy rapid economical practical met hod.The good experiment results with distinct image,reliable stability and good repeatability may be obtained using the method in RAPD amplification.