Multi#ex—CAPS技术及其在番茄遗传鉴定中的应用

 ：应用Multiplex—PCR和CAPS(Cleaved amplified polymorphic sequence)原理研究建立了Mutiplex—CAPS技术。该技术包括：(1)在PCR扩增反应体系加入2对、3对或多对核苷酸引物，经30～40个扩增循环后得到每对引物的特异扩增片段；(2)应用同一种限制性内切酶消化这些不同引物经扩增后获得的特异片段；(3)琼脂糖凝胶电泳溴化乙锭染色检测酶切片段长度的多态性。利用该技术对番茄(Lycopersicon es—cdentum)TA517及其近等基因系的分析表明，Multiplex—CAPS能同时检测2个、3个或多个CAPS标记的多态性，其效果等同于每个CAPS标记的多态性的叠加，重复性好，分析效率高，降低成本数倍。

The Establishment of Multiplex-CAPS Technique and Its Application for Genetic Characterization in Tomato

An efficient and reliable technique Multiplex- CAPS was developed for genetic characterization by combination of two principles Multi-PCR and CAPS (Cleaved amplified polymorphic sequence) in tomato. This technique included (1) using 2 or 3 or more than 3 primer pairs in the same amplification reaction, after 30-40 thermal cycles the PCR yield different products which correspond to each individual primer pair, (2) using only one enzyme to digest these different PCR products, (3) Using agrose gel electrophoresis to detect polymorphisms followed by ethidium staining. By comparison with CAPS, Multiplex - CAPS can detect the polymorphism of 2 or 3 or more than 3 primer pairs in the same amplification reaction at the same time. The polymorphism of Multiplex -CAPS equal exactly the polymorphism superimpose of each individual primer pair. Multiplex - CAPS has good re-producibility with high analytical effect, low cost, low labor and time consuming.