小麦叶片cDNA-AFLP扩增反应体系的优化

Optimization of cDNA-AFLP Amplification Reaction System in Wheat Leaves

【Objective】To establish and optimize cDNA-AFLP amplification reaction system for wheat leaves,cDNA-AFLP were analyzed,drought resistant gene were found in Xinchun 6.【Method】In the pot experiment of wheat at seedling stage under water stress,the optimization of cDNA-AFLP reaction system was studied by using wheat leaves as samples.Different levels of concentration of primer,dNTP mixture, Mg2+ and TaqDNA polymerase were studied in 20 μL amplification reaction system.【Result】Results showed that the quite unmber of clear and distinguishable TDFs bands could be found when the volume of primer was1.0 μL(40 mM),dNTP was 1.8 μL(2.5mM),Mg2+ was 2.2 μL(25 mM),TaqDNA was 0.5 μL(5U) in the preliminary amplification reaction system and the volume of primer was 0.6 μL(40mM),dNTP was2.0 μL(2.5 mM),Mg2+ was 1.6 μL(25 mM),TaqDNA was 0.4 μL(5U) in the selective amplificaion reaction system.【Conclusion】The study will set good base for segregating and cloning wheat gene to use cDNA-AFLP.