新疆野苹果S基因特异性PCR体系优化  

Optimization on PCR Amplification Procedure of the Malus sieversii S- gene

作　　者：王云[1] 秦伟[1] 王永丰[1] 

机构地区：[1]新疆农业大学特色果树研究中心,乌鲁木齐830052

出　　处：《新疆农业科学》2013年第11期2023-2030,共8页Xinjiang Agricultural Sciences

基　　金：新疆维吾尔自治区自然科学基金(2011211B15);国家自然科学基金(31200505);新疆维吾尔自治区果树重点学科

摘　　要：【目的】构建适合新疆野苹果自交不亲和基因的PCR扩增程序的优化体系。【方法】采用单因素比较试验,对影响S-RNase基因PCR反应体系的6个主要因素进行梯度扩增。【结果】在20μL的PCR总反应体系中,Mg2+的最适浓度为2.5 mmol/L,dNTP的最适浓度为0.2 mmol/L,引物的最适浓度为0.5 mmol/L,TaqDNA聚合酶的最适用量为0.75 U,模板DNA的最适用量为800 ng,最适退火温度为49℃。【结论】利用优化的扩增体系在99个新疆野苹果都获得了有效扩增,证明该体系为进行新疆野苹果S基因PCR扩增的最佳体系,具有良好的稳定性。[ Objective ] In order to construct a optimized system of PCR amplification procedure which was fit for Malus sieversii self- incompatibility gene （S - gcne）. [ Method ] The method of single factor comparison experiment was used to check out the effects of S - gene PCR reaction system of seven major factors in gradient amplification in this project. [ Result] The results showed that the optimized PCR system in 20 μL content contained 2.5 mmol/L Mg2＋ ,1 x Buffer,0.2 mmol/L dNTP,0.5 DNA template ,and the optimal annealing temperature was 49℃. optimal amplification system had effective amplification effect in mmol/L primer, 1U Taq polymerase, 800 ng [ Conclusion ] The studies suggested that the 99 samples of Malus sieversii, which proved that the system for Malus sieversii S - gene PCR amplification system was the best system with a fine stability.

关 键 词：新疆野苹果 S基因 PCR体系优化 

分 类 号：S188.1[农业科学—农业基础科学] S661.1