建立番木瓜性别研究的cDNA-AFLP分析体系

Construction of cDNA-AFLP Analysis System for Carica papaya Sex

以番木瓜幼苗为材料,利用多重PCR技术快速鉴定其性别,建立适合番木瓜性别研究的cDNA-AFLP分析体系,分析影响cDNA-AFLP分析体系的各种可能因素(RNA提取、酶切与连接效率、预扩增效果等),并对影响选择性扩增反应的各因子进行优化。结果表明,选择性扩增的最佳体系,即在反应体系中,各因素终浓度分别为:Tap酶0.06U/μL;Mg2+2.4mmol/L;dNTP2.0mmol/L;引物0.48μmol/L。通过优化体系得到的选择性扩增产物,银染检测后条带清晰稳定。表明成功建立了适合番木瓜性别研究的cDNA-AFLP分析体系。

Three sex types of papaya seedlings used as materials were determined by ways of multiple PRC. The factors influencing cDNA-AFLP (amplified fragment length polymorphism) analysis system were analyzed, such as RNA extraction, double enzymes restriction, adapter ligation and pre-amplification, etc., and an optimal selective amplification reaction system was established by optimizing the influencing factors, and their final concentration in the reaction system was 0.06 U/L for Tapase, 2.4 mmol/L for Mg2+, 2.0 ...