Abstract:

TRAP markers is widely used for its simplicity, repeatability and high efficiency, however, the technology is scarcely used in the study of tea plant, and tea tree TRAP-PCR system optimization has not been reported. Orthogonal design was applied for optimizing five factors (Mg2+, Taq DNA polymerase, dNTP, fixed primer, arbitrary primer) in the TRAP-PCR amplification system at four levels respectively. The best level of each factor were selected, a suitable TRAP-PCR reaction system was established, namely 20 μL reaction system containing 2μL 10×PCR Buffer(Mg2+ free)，50 ng DNA，1.75 mmo/L Mg2+，0.50 U Taq DNA polymerase, 0.20 mmol/L dNTP, 0.20 mmol/L arbitrary primer and 0.2 mmol/L fixed primer. TRAP polymorphism was studied using sixteen tea germplasm.