Optimal reaction system for random amplified polymorphic DNA with endangered Emmenopterys henryi

Key words: 
• botany /
• Emmenopterys henryi(Henry emmenoterys) /
• genomic DNA /
• randomamplified polymorphic DNA(RAPD)

Abstract: Genome DNA from leaves of the Emmenopterys henryi tree(Henry emmenopterys) in the Shennongjia area of Hubei Province, China, was extracted to optimize its random amplified polymorphic DNA(RAPD) reaction conditions, which included concentrations of DNA polymerase, primers, Mg2+,dNTP, and DNA.Resultsindicated that genome DNA of E. henryi had good amplification with the following conditions: 25L solutionwith 1.33 10- 3 katL- 1 Taq DNA polymerase, 0.5molL- 1 random primer, 2.6 mmolL- 1 Mg2+,220molL- 1 dNTP, and 4.40 mgL- 1 genomic DNA. The polymerase chain reaction(PCR) amplification programwas as follows: predenaturing at 94℃ for5 min, followed by 40 cycles of denaturing at 94℃ for1 min,annealing at 43℃ for 1 min, extension at 72℃ for 2 min, and final extension at 72℃ for 8 min. Such a system and reaction programs could achieve comparatively stable amplification.[Ch, 6 fig. 1 tab. 8 ref.]