关键词: 小蓬竹, RAPDPCR, 引物, dNTPs, Taq DNA聚合酶

Abstract: The Drepanostachyum luodianense RAPDPCR reaction system was optimized for the analysis of genetic variation and structure of D. luodianense. The genomic DNA of D. luodianense was extracted by improved CTAB method. Then， both single factor test and orthogonal design were applied to study the effects of main factors on the RAPDPCR system for D. luodianense, in which the main factors included the concentration of dNTPs， primers and Mg2+. The content of template DNA and Taq DNA polymerase and an optimal 20 μL RAPDPCR reaction system for D. luodianense was established， including 1/10 volume 10×PCR buffer， 100 μmol·L-1 dNTPs， 30 ng template DNA， 10 U Taq DNA polymerase， 02 μmol·L-1 primers and 150 mmol·L-1 Mg2+. The optimized reaction program was initially at 94℃ for 5 min， followed by 35 cycles at 94℃ for 1 min， at 35℃ for 30 s， at 72℃ for 90 s， and then held at 72℃ for 7 min， and finally kept at 4℃．

Key words: Drepanostachyum luodianense, RAPDPCR, primers, dNTPs, Taq DNA polymerase